Glycan Analysis – VectorLabs https://staging.vectorlabs.com From linker design and synthesis, bioconjugated design and manufacturing, biomolecule labeling and functionalization, and detection system and solutions, our team is here to help you move forward, with impact. Wed, 22 Oct 2025 17:14:50 +0000 en-US hourly 1 https://wordpress.org/?v=6.8.3 https://staging.vectorlabs.com/wp-content/uploads/2024/11/V-favicon-02-100x100.png Glycan Analysis – VectorLabs https://staging.vectorlabs.com 32 32 Glysite™ Explorer in situ PLA Glycan Detection Kit & Glysite™ Explorer Lectins https://staging.vectorlabs.com/products/glysite-explorer-lectins/ Wed, 26 Mar 2025 02:33:26 +0000 https://staging.vectorlabs.com/products/gek-1000/ The Glysite™ Explorer in situ PLA Glycan Detection Kit is a fully integrated tool for the spatial detection of glycans proximal to a protein of interest in FFPE tissues, FFPE cell pellets and fixed cells. This system integrates a curated panel of Vector’s Glysite™ Explorer lectins with Navinci’s proprietary in situ Proximity Ligation Assay (isPLA). Researchers can flexibly select specific Glysite™ Explorer lectins covering the major categories of glycan types to pair with a primary antibody targeting their protein of interest. This provides a simple approach to understand glycan-protein proximity with a spatial context.  

Specification

Glysite™ Explorer in situ PLA Glycan Detection Kit

Unit Size 1 kit
Applications Immunohistochemistry, Glycobiology
Sample Type FFPE Tissue, FFPE Cell Pellet, Fixed Cells
Recommended Storage Store reagents in original bottles at 4°C and -20°C accordingly

Glysite™ Explorer Lectins

Unit Size 0.1 mL per lectin
Applications Immunohistochemistry, Glycobiology
Sample Type FFPE Tissues, FFPE Cell Pellets, Fixed Cells
Recommended Storage Store reagents in original bottles at 4°C. Do not freeze.
Recommended Usage Prior to use, 50x Lectin should be diluted to 1x with Protein Diluent.
Sugar Specificity
  • WGA Lectin: Terminal GlcNAcβ, Terminal GlcNAcα, Terminal N-acetyl-containing glycans
  • LCA Lectin: α1-6 fucose
  • PHA-L Lectin: β1-6 branched N-glycans, binds tri- and tetraantennary
  • ECL Lectin: Terminal type 2 LacNAc, Terminal type 2 LacdiNAc
  • Jacalin Lectin: Core 1 and 3 O-glycans, 3-substituted GalNAcα
  • GNL Lectin: Terminal α1-3 or α1-6 mannose
  • MAL II Lectin: α2-3-sialylated Galβ1-3GalNAc in O-glycans, 3′ sulfated Galβ
  • SNA Lectin: α2-6 sialylated LacNAc, α2-6 sialylated LacdiNAc
  • WFA Lectin: Terminal GalNAcβ, Terminal GalNAcα, Terminal multiantennary LacNAc
  • AAL Lectin: α-Fucose

Glysite™ Explorer in situ PLA Glycan Detection Kit (GEK-1000) contains the following reagents:

Box 1 Reagents (4°C storage, do not freeze):

  • BLOXALL® Blocking Solution
  • Blocking Buffer 1
  • Blocking Buffer 2A
  • Blocking Buffer 2B (25x)
  • Protein Diluent
  • Lectin Probe (50x)
  • Mouse Probe (50x)
  • Rabbit Probe (50x)
  • ImmPRESS® HRP Reagent (400x)
  • ImmPRESS® HRP Diluent
  • ImmPACT® DAB Reagent (33x)
  • ImmPACT® DAB Diluent
  • Hematoxylin QS

Box 2 Reagents (-20°C storage):

  • Buffer 1 (5x)
  • Enzyme 1 (40x)
  • Buffer 2 (5x)
  • Enzyme 2 (40x)

Technical Information

The Glysite™ Explorer in situ PLA Glycan Detection Kit leverages Vector’s well-known immunohistochemistry portfolio and lectins with Navinci’s proven isPLA technology. This assay enables researchers to detect glycosylation proximal to specific proteins in a spatial context.

Simplified scheme of Glysite™ Explorer in situ PLA Glycan Detection Kit. A. Lectin binds to target glycan and primary antibody binds to target protein. B. Oligo probes bind to primary antibody and lectin. C. When oligo probes are in proximity (~40nm), ligation occurs. D. Amplification and detection produces a colored precipitate at the site of proximity. Recommended experiments before using the Glysite™ Explorer in situ PLA Glycan Detection Kit:

  • Perform standard Immunohistochemistry (IHC), Immunofluorescence (IF), or other assays to characterize expression/abundance of protein of interest and ensure the primary antibody is at the optimal concentration and shows correct specificity with great signal-to-noise ratios.
  • Perform standard Immunohistochemistry (IHC), Immunofluorescence (IF) or other assays to confirm the expression/abundance of the glycan of interest. Vector Laboratories offers (GSK-1000, GSK-2000, GSK-3000). These kits are fully integrated kits for the detection of glycan expression in fixed cells and tissue sections.

The post Glysite™ Explorer <i>in situ</i> PLA Glycan Detection Kit & Glysite™ Explorer Lectins appeared first on VectorLabs.

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Description

The Glysite™ Explorer in situ PLA Glycan Detection Kit is a fully integrated tool for the spatial detection of glycans proximal to a protein of interest in FFPE tissues, FFPE cell pellets and fixed cells. This system integrates a curated panel of Vector’s Glysite™ Explorer Lectins with Navinci’s proprietary in situ Proximity Ligation Assay (isPLA). Researchers can flexibly select specific Glysite™ Explorer Lectins covering the major categories of glycan types to pair with a primary antibody targeting their protein of interest. This provides a simple approach to understand glycan-protein proximity with a spatial context.  

Specification

Glysite™ Explorer in situ PLA Glycan Detection Kit

Unit Size1 kit
ApplicationsImmunohistochemistry, Glycobiology
Sample TypeFFPE Tissue, FFPE Cell Pellet, Fixed Cells
Recommended StorageStore reagents in original bottles at 4°C and -20°C accordingly

Glysite™ Explorer Lectins

Unit Size0.1 mL per lectin
ApplicationsImmunohistochemistry, Glycobiology
Sample TypeFFPE Tissues, FFPE Cell Pellets, Fixed Cells
Recommended StorageStore reagents in original bottles at 4°C. Do not freeze.
Recommended UsageEach Glysite Explorer Lectin is sufficient to perform 50 reactions when using 100uL per reaction. Prior to use, Explorer Lectin (50x) should be diluted to 1x with Protein Diluent.
Sugar Specificity
  • WGA Lectin: Terminal GlcNAcβ, Terminal GlcNAcα, Terminal N-acetyl-containing glycans
  • LCA Lectin: α1-6 fucose
  • PHA-L Lectin: β1-6 branched N-glycans, binds tri- and tetraantennary
  • ECL Lectin: Terminal type 2 LacNAc, Terminal type 2 LacdiNAc
  • Jacalin Lectin: Core 1 and 3 O-glycans, 3-substituted GalNAcα
  • GNL Lectin: Terminal α1-3 or α1-6 mannose
  • MAL II Lectin: α2-3-sialylated Galβ1-3GalNAc in O-glycans, 3′ sulfated Galβ
  • SNA Lectin: α2-6 sialylated LacNAc, α2-6 sialylated LacdiNAc
  • WFA Lectin: Terminal GalNAcβ, Terminal GalNAcα, Terminal multiantennary LacNAc
  • AAL Lectin: α-Fucose

Kit Components

Glysite™ Explorer in situ PLA Glycan Detection Kit (GEK-1000) contains the following reagents:

Box 1 Reagents (4°C storage, do not freeze):

  • BLOXALL® Blocking Solution
  • Blocking Buffer 1
  • Blocking Buffer 2A
  • Blocking Buffer 2B (25x)
  • Protein Diluent
  • Lectin Probe (50x)
  • Mouse Probe (50x)
  • Rabbit Probe (50x)
  • ImmPRESS® HRP Reagent (400x)
  • ImmPRESS® HRP Diluent
  • ImmPACT® DAB Reagent (33x)
  • ImmPACT® DAB Diluent
  • Hematoxylin QS

Box 2 Reagents (-20°C storage):

  • Buffer 1 (5x)
  • Enzyme 1 (40x)
  • Buffer 2 (5x)
  • Enzyme 2 (40x)

Documents

FAQs

The Glysite Explorer Kit allows the visualization of a glycan and a protein when they are within 40nm of each other.

Customers will need to purchase a Glysite Explorer Lectin, and for additional reagent requirements, please see the User Guide.

  • The Glysite Explorer kit has been validated on human specimens.
  • Other species of tissues should also be compatible, but the primary antibody species should be selected to avoid endogenous IgG detection.

The Glysite Explorer Kit has been validated on formalin-fixed, paraffin-embedded (FFPE) tissues, FFPE cells, and fixed cells. The kit is not recommended for use on frozen tissue specimens at this time.

  • The concentrations and incubation times in the Glysite Explorer Kit have been optimized for 4-6 µm sections. We have not evaluated the performance of this kit on thick sections or free-floating sections.
  • For users who want to stain thick sections, the protocol will need to be modified.

  • The Glysite Explorer Kit includes two separate secondary antibody reagents: one anti-mouse and one anti-rabbit. This allows users to pair either a mouse or a rabbit primary antibody with their chosen Glysite Explorer Lectin, depending on the needs of the experiment.
  • We do not recommend the use of a primary antibody from the same species as the tissue being studied—for example, do not use a mouse primary on mouse tissue—to minimize nonspecific binding and reduce the risk of false-positive signals from endogenous IgG.

  • Multiplexing with the Glysite Explorer Kit is generally not recommended.
  • This kit has not been validated to perform multiplexing. In some cases, it may be possible if detection reagents do not cross-react and the proper controls are included.

  • Positive controls
    • Characterize the expression of the protein of interest in your specimen using standard Immunohistochemistry (IHC), Immunofluorescence (IF) or other workflows.
      • Determine the optimal dilution for the primary antibody in a standard IHC assay as this will be the recommended starting dilution for the isPLA assay.
    • Characterize the expression of the glycan of interest in your specimen using standard Immunohistochemistry (IHC), Immunofluorescence (IF) or other workflows.
  •  Negative control:
    • Omit the Glysite Explorer Lectin and/or primary antibody from the workflow to demonstrate whether non-specific binding or background staining is present.

Reagent volumes should be determined by the researcher to align with the dimensions of their specimens. Volumes sufficient to cover the specimens should be applied, and samples should not be allowed to dry out during the incubation steps.

The number of specimens that can be stained will vary depending on specimen size. The Glysite Explorer Kit provides 5 mL of working solution. If using 50-100µL of reagent per slide, there is sufficient volume to stain 50-100 specimens.

Slides stained with the Glysite Explorer Kit can be mounted with either an aqueous or a non-aqueous mounting medium. For non-aqueous mounting, we recommend using VectaMount® PT Permanent Mounting Medium (H-5600-60) or VectaMount® Express Mounting Medium (H-5700-60). For aqueous mounting, we recommend using VectaMount® AQ Aqueous Mounting Medium (H-5501-60).

  • The Glysite Explorer Kit generates distinct, round signals called puncta at the site of proximity. These are best visualized using at least a 20x objective, and sometimes a 40x may be required, especially when epitope density is low.
  • If the counterstain is too dark, it can obscure isPLA signals. Hematoxylin QS incubation should be optimized for each specimen to ensure positive signals are not masked. Even 1 second of staining is often sufficient, if performed at all.
  • The overall appearance will vary by specimen and proximal target abundance.

The average run-time will depend on the duration of the primary antibody incubation. If your primary antibody incubation is 30 minutes, the total run time is approximately 8 hours.

If the assay cannot be completed within a single day, the tissue sections can be stored overnight at 4°C in Protein Diluent after incubation of the Lectin Solution in step 13. Do not allow the sample to dry out.

The kit is supplied with ImmPACT DAB, Vector’s most sensitive peroxidase substrate. Other peroxidase substrates can be substituted, and the Glysite Explorer Kit has been validated with ImmPACT® NovaRed (Catalog Number: SK-4805), ImmPACT® VIP (SK-4605), and ImmPACT® SG (SK-4705). However, not all peroxidase substrates are equivalent in sensitivity and staining quality and should be optimized to achieve desired results.

No, the Glysite Explorer Kit is not currently available with a fluorescent readout.

  • The Glysite Explorer Kit is supplied in two boxes, and the labels indicate the proper storage conditions. Box 1 should be stored at 4°C. Box 2 should be stored at -20°C.
  • The Glysite Explorer Lectins should be stored at 4°C.

Technical Information

The Glysite™ Explorer in situ PLA Glycan Detection Kit leverages Vector’s well-known immunohistochemistry portfolio and lectins with Navinci’s proven isPLA technology. This assay enables researchers to detect glycosylation proximal to specific proteins in a spatial context.

workflow icons

A

B

C

D

Simplified scheme of Glysite™ Explorer in situ PLA Glycan Detection Kit. A. Lectin binds to target glycan and primary antibody binds to target protein. B. Oligo probes bind to primary antibody and lectin. C. When oligo probes are in proximity (~40nm), ligation occurs. D. Amplification and detection produces a colored precipitate at the site of proximity.

Recommended experiments before using the Glysite™ Explorer in situ PLA Glycan Detection Kit:

  • Perform standard Immunohistochemistry (IHC), Immunofluorescence (IF), or other assays to characterize expression/abundance of protein of interest and ensure the primary antibody is at the optimal concentration and shows correct specificity with great signal-to-noise ratios.
  • Perform standard Immunohistochemistry (IHC), Immunofluorescence (IF) or other assays to confirm the expression/abundance of the glycan of interest. Vector Laboratories offers (GSK-1000, GSK-2000, GSK-3000). These kits are fully integrated kits for the detection of glycan expression in fixed cells and tissue sections.

The post Glysite™ Explorer <i>in situ</i> PLA Glycan Detection Kit & Glysite™ Explorer Lectins appeared first on VectorLabs.

]]> Ricin A Chain https://staging.vectorlabs.com/products/ricin-a-chain/ Sun, 10 Dec 2023 16:05:50 +0000 https://staging.vectorlabs.com/?post_type=product&p=32833 Description

Ricin A chain has been isolated from toxic RCA II. Our ricin A chain has less than 0.01% of the toxicity of the native lectin in a cell culture test system, yet is as potent as native ricin in a cell-free protein synthesis assay. The A chain can be linked to other proteins such as antibodies. These antibody-hybrid molecules can be used to kill specific cells with the appropriate antigenic determinant exposed, such as tumor markers on surfaces of malignant cells. The A chain is supplied in buffered saline with sodium azide as a preservative.

Specifications

Unit Size 0.5 mg
Applications Cellular toxicity
Recommended Storage
2-8 °C
Solution
10 mM phosphate, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 50 mM 2-mercaptoethanol, 0.1% polyethylene glycol (M.W. 3300)
Concentration
1 mg/mL
Conjugate
Unconjugated

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Description

Ricin A chain has been isolated from toxic RCA II. Our ricin A chain has less than 0.01% of the toxicity of the native lectin in a cell culture test system, yet is as potent as native ricin in a cell-free protein synthesis assay. The A chain can be linked to other proteins such as antibodies. These antibody-hybrid molecules can be used to kill specific cells with the appropriate antigenic determinant exposed, such as tumor markers on surfaces of malignant cells. The A chain is supplied in buffered saline with sodium azide as a preservative.

Specifications

Unit Size0.5 mg
ApplicationsCellular toxicity
Recommended Storage
2-8 °C
Solution
10 mM phosphate, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 50 mM 2-mercaptoethanol, 0.1% polyethylene glycol (M.W. 3300)
Concentration
1 mg/mL
Conjugate
Unconjugated

Documents

The post Ricin A Chain appeared first on VectorLabs.

]]> Ricin B Chain https://staging.vectorlabs.com/products/ricin-b-chain/ Sun, 10 Dec 2023 16:05:40 +0000 https://staging.vectorlabs.com/?post_type=product&p=32839 Description

Ricin B chain has been isolated from highly purified RCA II. B chain is the component of ricin which binds surface carbohydrate receptors and facilitates the transport of ricin into the cell. Although our ricin B chain has a substantially lower toxicity than native ricin in both cell culture test systems and cell-free protein synthesis assays, essentially all of the galactose/N-acetylgalactosamine binding activity is retained.

Specifications

Unit Size 0.5 mg
Applications Glycoprotein binding assays
Recommended Storage
2-8 °C
Solution
10 mM phosphate, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 50 mM 2-mercaptoethanol
Concentration
1 mg/mL
Conjugate
Unconjugated

The post Ricin B Chain appeared first on VectorLabs.

]]>

Description

Ricin B chain has been isolated from highly purified RCA II. B chain is the component of ricin which binds surface carbohydrate receptors and facilitates the transport of ricin into the cell. Although our ricin B chain has a substantially lower toxicity than native ricin in both cell culture test systems and cell-free protein synthesis assays, essentially all of the galactose/N-acetylgalactosamine binding activity is retained.

Specifications

Unit Size0.5 mg
ApplicationsGlycoprotein binding assays
Recommended Storage
2-8 °C
Solution
10 mM phosphate, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 50 mM 2-mercaptoethanol
Concentration
1 mg/mL
Conjugate
Unconjugated

Documents

The post Ricin B Chain appeared first on VectorLabs.

]]> Click-&-Go® IsoTaG Kit for Intact Glycopeptides Profiling https://staging.vectorlabs.com/products/click-go-isotag-kit-for-intact-glycopeptides-profiling-2/ Tue, 19 Sep 2023 19:51:21 +0000 https://staging.vectorlabs.com/?post_type=product&p=22185 Specifications
Enrichment target Azide-modified proteins
Enrichment Technology Biotin-streptavidin based enrichment
Number of Enrichments 25
Storage Conditions 4C
Shipping Conditions Ambient temperature

Description

While there has been much interest in profiling the intact glycoproteome, the complexity of glycoproteoforms (and more broadly, all proteoforms) remains challenging to completely define. Mass spectrometry (MS) is commonly employed for characterization of complex proteomic samples. A popular strategy for protein identification is the bottom-up shotgun proteomics approach. In this method, a mixture of proteins is subjected to proteolytic digestion, the resulting peptides are separated by LC and detected by MS, and their parent proteins are inferred from the assigned peptide sequences.

To convert MS data acquired from proteolytic digests into protein identifications, tandem MS can be used to obtain sequence information for individual peptides, followed by comparing an in-silico proteolytic digest of an organism’s proteome. Typically, only the most abundant peptides are selected for fragmentation (Figure 2), whereas data for those peptides in relatively low quantities are not obtained. An inherent problem in shotgun proteomics is identifying proteins of low abundance, such as biomarkers for disease states, against a background of proteins whose concentrations can span up to 12 orders of magnitude.

Figure 1. Metabolic labeling with a chemically functionalized glycan, chemical tagging and enrichment using an isotopic recoding affinity probe

To address the unique challenges of the global characterization of the intact glycoproteome, a mass-independent chemical glycoproteomics platform, termed isotope targeted glycoproteomics (IsoTag) was developed by the Carolyn Bertozzi group. The platform is comprised of four central components: (i) metabolic labeling with a chemically functionalized glycan, (ii) chemical tagging and enrichment using an isotopic recoding affinity probe, (iii) directed tandem MS, and (iv) targeted glycopeptide assignment (Figure 2).

Figure 2. Traditional proteomics and Iso-Tag-directed proteomics workflow

IsoTaG is performed by isotopic recoding and enrichment of metabolically labeled glycoproteins followed by directed tandem MS (MS2 or MSn) analysis and intact glycopeptide assignment. Isotopic recoding is accomplished by metabolic labeling of cell or tissue samples with azide- or alkyne-functionalized sugars, followed by chemical conjugation with a biotin probe bearing a unique isotopic signature.

Some examples of sugar labels are peracetylated N-azidoacetylmannosamine (Ac4ManNAz), which is converted to the corresponding azidosialic acid (SiaNAz), and peracetylated N-azidoacetylgalactosamine (Ac4GalNAz), which is metabolized to label glycans possessing N-acetylglucosamine (GlcNAc) or N-acetylgalactosamine (GalNAc) (not provided with kit).

In order to perform isotopic tagging, the kit provides two cleavable IsoTaG probes encoded by zero [M] and two [M + 2] deuterium atoms. Probes with different encoding can be provided by Click Chemistry Tools though custom synthesis. The IsoTaG probe with zero, and that with two deuterium atoms [M, M + 2] can be used in different proportions; 1:1, 1:2, 1:3 and 1:4. Pattern recognition with isotopic ratio of 1:3 showed the highest fidelity.

Figure 3. Cleavable IsoTaG probe encoded by zero deuterium atoms [M] (R = H) and two deuterium atoms [M+2] (R = D)

Through these probes, a unique isotopic signature is embedded exclusively into the glycopeptides. The isotopic signature serves as a computationally recognizable full-scan MS reporter. A computational algorithm, termed isotopic signature transfer and mass pattern prediction (IsoStamp), for the detection of recorded species in full-scan mass spectra, was also developed by the Carolyn Bertozzi group. IsoStamp compares observed and predicted isotopic envelopes to identify chemically tagged species in full-scan mass spectra.

IsoTag has the potential to enhance any proteomics platform that employs chemical labeling for targeted protein identification, including isotope-coded affinity tagging, isobaric tagging for relative and absolute quantitation, and chemical tagging strategies for post-translational modification.

[caption id="attachment_26280" align="alignnone" width="380"] Schematic Workflow[/caption]

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]]>
IsoTaG is a mass-independent chemical glycoproteomics platform that helps to address unique challenges of the global characterization of the intact glycoproteome. In this approach a unique isotopic signature is embedded exclusively into the glycopeptides and it serves as a computationally recognizable full-scan MS reporter. A computational algorithm, termed isotopic signature transfer and mass pattern prediction (IsoStamp), for the detection of recoded species in full-scan mass spectra, was also developed by the Carolyn Bertozzi group.

Description

While there has been much interest in profiling the intact glycoproteome, the complexity of glycoproteoforms (and more broadly, all proteoforms) remains challenging to completely define. Mass spectrometry (MS) is commonly employed for characterization of complex proteomic samples. A popular strategy for protein identification is the bottom-up shotgun proteomics approach. In this method, a mixture of proteins is subjected to proteolytic digestion, the resulting peptides are separated by LC and detected by MS, and their parent proteins are inferred from the assigned peptide sequences.

To convert MS data acquired from proteolytic digests into protein identifications, tandem MS can be used to obtain sequence information for individual peptides, followed by comparing an in-silico proteolytic digest of an organism’s proteome. Typically, only the most abundant peptides are selected for fragmentation (Figure 2), whereas data for those peptides in relatively low quantities are not obtained. An inherent problem in shotgun proteomics is identifying proteins of low abundance, such as biomarkers for disease states, against a background of proteins whose concentrations can span up to 12 orders of magnitude.

iso tag img one
Figure 1. Metabolic labeling with a chemically functionalized glycan, chemical tagging and enrichment using an isotopic recoding affinity probe

To address the unique challenges of the global characterization of the intact glycoproteome, a mass-independent chemical glycoproteomics platform, termed isotope targeted glycoproteomics (IsoTag) was developed by the Carolyn Bertozzi group. The platform is comprised of four central components: (i) metabolic labeling with a chemically functionalized glycan, (ii) chemical tagging and enrichment using an isotopic recoding affinity probe, (iii) directed tandem MS, and (iv) targeted glycopeptide assignment (Figure 2).

iso tag img two
Figure 2. Traditional proteomics and Iso-Tag-directed proteomics workflow

IsoTaG is performed by isotopic recoding and enrichment of metabolically labeled glycoproteins followed by directed tandem MS (MS2 or MSn) analysis and intact glycopeptide assignment. Isotopic recoding is accomplished by metabolic labeling of cell or tissue samples with azide- or alkyne-functionalized sugars, followed by chemical conjugation with a biotin probe bearing a unique isotopic signature.

Some examples of sugar labels are peracetylated N-azidoacetylmannosamine (Ac4ManNAz), which is converted to the corresponding azidosialic acid (SiaNAz), and peracetylated N-azidoacetylgalactosamine (Ac4GalNAz), which is metabolized to label glycans possessing N-acetylglucosamine (GlcNAc) or N-acetylgalactosamine (GalNAc) (not provided with kit).

In order to perform isotopic tagging, the kit provides two cleavable IsoTaG probes encoded by zero [M] and two [M + 2] deuterium atoms. Probes with different encoding can be provided by Click Chemistry Tools though custom synthesis. The IsoTaG probe with zero, and that with two deuterium atoms [M, M + 2] can be used in different proportions; 1:1, 1:2, 1:3 and 1:4. Pattern recognition with isotopic ratio of 1:3 showed the highest fidelity.

iso tag img three
Figure 3. Cleavable IsoTaG probe encoded by zero deuterium atoms [M] (R = H) and two deuterium atoms [M+2] (R = D)

Through these probes, a unique isotopic signature is embedded exclusively into the glycopeptides. The isotopic signature serves as a computationally recognizable full-scan MS reporter. A computational algorithm, termed isotopic signature transfer and mass pattern prediction (IsoStamp), for the detection of recorded species in full-scan mass spectra, was also developed by the Carolyn Bertozzi group. IsoStamp compares observed and predicted isotopic envelopes to identify chemically tagged species in full-scan mass spectra.

IsoTag has the potential to enhance any proteomics platform that employs chemical labeling for targeted protein identification, including isotope-coded affinity tagging, isobaric tagging for relative and absolute quantitation, and chemical tagging strategies for post-translational modification.

Specifications

Enrichment targetAzide-modified proteins
Enrichment TechnologyBiotin-streptavidin based enrichment
Number of Enrichments25
Storage Conditions4C
Shipping ConditionsAmbient temperature

Schematic Workflow

Biotinylation reagents 1442

Documents

Selected References

  1. Woo, C.M., et al. (2017). Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O‑Glycopeptides from Whole Cell Proteomess. J. Proteome Res., 16: 1706−18. [PubMed]
  2. Woo, C.M., et al. (2017). Mapping and Quantification of Over 2000 O-linked Glycopeptides in Activated Human T Cells with Isotope-Targeted Glycoproteomics (Isotag). Mol. Cell. Proteomics 17: 764-75. [PubMed]
  3. Goa, G., et al. (2017). Small Molecule Interactome Mapping by Photoaffinity Labeling Reveals Binding Site Hotspots for the NSAIDs. J. Am. Chem. Soc., 140: 4259−68. [PubMed]
  4. Woo, C.M., et al. (2015). Isotope-targeted glycoproteomics (IsoTaG): a mass-independent platform for intact N- and O-glycopeptide discovery and analysis.Nat Methods., 12: 561-7. [PubMed]
  5. Weerapana, E., et al. (2010). Quantitative reactivity profiling predicts functional cysteines in proteomes. Nature 468: 790−5. [PubMed]

IsoStamp v2

Download
zip archive

The post Click-&-Go® IsoTaG Kit for Intact Glycopeptides Profiling appeared first on VectorLabs.

]]> Click-&-Go® IsoTaG Kit for Intact Glycopeptides Profiling https://staging.vectorlabs.com/products/click-go-isotag-kit-for-intact-glycopeptides-profiling/ Tue, 19 Sep 2023 19:50:52 +0000 https://staging.vectorlabs.com/?post_type=product&p=22174 Description

While there has been much interest in profiling the intact glycoproteome, the complexity of glycoproteoforms (and more broadly, all proteoforms) remains challenging to completely define. Mass spectrometry (MS) is commonly employed for characterization of complex proteomic samples. A popular strategy for protein identification is the bottom-up shotgun proteomics approach. In this method, a mixture of proteins is subjected to proteolytic digestion, the resulting peptides are separated by LC and detected by MS, and their parent proteins are inferred from the assigned peptide sequences.

To convert MS data acquired from proteolytic digests into protein identifications, tandem MS can be used to obtain sequence information for individual peptides, followed by comparing an in-silico proteolytic digest of an organism’s proteome. Typically, only the most abundant peptides are selected for fragmentation, whereas data for those peptides in relatively low quantities are not obtained. An inherent problem in shotgun proteomics is identifying proteins of low abundance, such as biomarkers for disease states, against a background of proteins whose concentrations can span up to 12 orders of magnitude.

To address the unique challenges of the global characterization of the intact glycoproteome, a mass-independent chemical glycoproteomics platform, termed isotope targeted glycoproteomics (IsoTag) was developed by the Carolyn Bertozzi group. The platform is comprised of four central components: (i) metabolic labeling with a chemically functionalized glycan, (ii) chemical tagging and enrichment using an isotopic recoding affinity probe, (iii) directed tandem MS, and (iv) targeted glycopeptide assignment.

IsoTaG is performed by isotopic recoding and enrichment of metabolically labeled glycoproteins followed by directed tandem MS (MS2 or MSn) analysis and intact glycopeptide assignment. Isotopic recoding is accomplished by metabolic labeling of cell or tissue samples with azide- or alkyne-functionalized sugars, followed by chemical conjugation with a biotin probe bearing a unique isotopic signature.

Some examples of sugar labels are peracetylated N-azidoacetylmannosamine (Ac4ManNAz), which is converted to the corresponding azidosialic acid (SiaNAz), and peracetylated N-azidoacetylgalactosamine (Ac4GalNAz), which is metabolized to label glycans possessing N-acetylglucosamine (GlcNAc) or N-acetylgalactosamine (GalNAc) (not provided with kit).

In order to perform isotopic tagging, the kit provides two cleavable IsoTaG probes encoded by zero [M] and two [M + 2] deuterium atoms. Probes with different encoding can be provided by Click Chemistry Tools though custom synthesis. The IsoTaG probe with zero, and that with two deuterium atoms [M, M + 2] can be used in different proportions; 1:1, 1:2, 1:3 and 1:4. Pattern recognition with isotopic ratio of 1:3 showed the highest fidelity.

Through these probes, a unique isotopic signature is embedded exclusively into the glycopeptides. The isotopic signature serves as a computationally recognizable full-scan MS reporter. A computational algorithm, termed isotopic signature transfer and mass pattern prediction (IsoStamp), for the detection of recorded species in full-scan mass spectra, was also developed by the Carolyn Bertozzi group. IsoStamp compares observed and predicted isotopic envelopes to identify chemically tagged species in full-scan mass spectra.

IsoTag has the potential to enhance any proteomics platform that employs chemical labeling for targeted protein identification, including isotope-coded affinity tagging, isobaric tagging for relative and absolute quantitation, and chemical tagging strategies for post-translational modification.

Specifications

Enrichment target Alkyne-modified proteins
Enrichment Technology Biotin-streptavidin based enrichment
Number of Enrichments 25
Storage Conditions 4C
Shipping Conditions Ambient temperature

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IsoTaG is a mass-independent chemical glycoproteomics platform that helps to address unique challenges of the global characterization of the intact glycoproteome. In this approach a unique isotopic signature is embedded exclusively into the glycopeptides and it serves as a computationally recognizable full-scan MS reporter. A computational algorithm, termed isotopic signature transfer and mass pattern prediction (IsoStamp), for the detection of recoded species in full-scan mass spectra, was also developed by the Carolyn Bertozzi group.

Description

While there has been much interest in profiling the intact glycoproteome, the complexity of glycoproteoforms (and more broadly, all proteoforms) remains challenging to completely define. Mass spectrometry (MS) is commonly employed for characterization of complex proteomic samples. A popular strategy for protein identification is the bottom-up shotgun proteomics approach. In this method, a mixture of proteins is subjected to proteolytic digestion, the resulting peptides are separated by LC and detected by MS, and their parent proteins are inferred from the assigned peptide sequences.

To convert MS data acquired from proteolytic digests into protein identifications, tandem MS can be used to obtain sequence information for individual peptides, followed by comparing an in-silico proteolytic digest of an organism’s proteome. Typically, only the most abundant peptides are selected for fragmentation (Figure 2), whereas data for those peptides in relatively low quantities are not obtained. An inherent problem in shotgun proteomics is identifying proteins of low abundance, such as biomarkers for disease states, against a background of proteins whose concentrations can span up to 12 orders of magnitude.

iso tag img one
Figure 1. Metabolic labeling with a chemically functionalized glycan, chemical tagging and enrichment using an isotopic recoding affinity probe

To address the unique challenges of the global characterization of the intact glycoproteome, a mass-independent chemical glycoproteomics platform, termed isotope targeted glycoproteomics (IsoTag) was developed by the Carolyn Bertozzi group. The platform is comprised of four central components: (i) metabolic labeling with a chemically functionalized glycan, (ii) chemical tagging and enrichment using an isotopic recoding affinity probe, (iii) directed tandem MS, and (iv) targeted glycopeptide assignment (Figure 2).

iso tag img two
Figure 2. Traditional proteomics and Iso-Tag-directed proteomics workflow

IsoTaG is performed by isotopic recoding and enrichment of metabolically labeled glycoproteins followed by directed tandem MS (MS2 or MSn) analysis and intact glycopeptide assignment. Isotopic recoding is accomplished by metabolic labeling of cell or tissue samples with azide- or alkyne-functionalized sugars, followed by chemical conjugation with a biotin probe bearing a unique isotopic signature.

Some examples of sugar labels are peracetylated N-azidoacetylmannosamine (Ac4ManNAz), which is converted to the corresponding azidosialic acid (SiaNAz), and peracetylated N-azidoacetylgalactosamine (Ac4GalNAz), which is metabolized to label glycans possessing N-acetylglucosamine (GlcNAc) or N-acetylgalactosamine (GalNAc) (not provided with kit).

In order to perform isotopic tagging, the kit provides two cleavable IsoTaG probes encoded by zero [M] and two [M + 2] deuterium atoms. Probes with different encoding can be provided by Click Chemistry Tools though custom synthesis. The IsoTaG probe with zero, and that with two deuterium atoms [M, M + 2] can be used in different proportions; 1:1, 1:2, 1:3 and 1:4. Pattern recognition with isotopic ratio of 1:3 showed the highest fidelity.

iso tag img three
Figure 3. Cleavable IsoTaG probe encoded by zero deuterium atoms [M] (R = H) and two deuterium atoms [M+2] (R = D)

Through these probes, a unique isotopic signature is embedded exclusively into the glycopeptides. The isotopic signature serves as a computationally recognizable full-scan MS reporter. A computational algorithm, termed isotopic signature transfer and mass pattern prediction (IsoStamp), for the detection of recorded species in full-scan mass spectra, was also developed by the Carolyn Bertozzi group. IsoStamp compares observed and predicted isotopic envelopes to identify chemically tagged species in full-scan mass spectra.

IsoTag has the potential to enhance any proteomics platform that employs chemical labeling for targeted protein identification, including isotope-coded affinity tagging, isobaric tagging for relative and absolute quantitation, and chemical tagging strategies for post-translational modification.

Specifications

Enrichment targetAlkyne-modified proteins
Enrichment TechnologyBiotin-streptavidin based enrichment
Number of Enrichments25
Storage Conditions4C
Shipping ConditionsAmbient temperature

Schematic Workflow

Biotinylation reagents 1442

Documents

Selected References

  1. Woo, C.M., et al. (2017). Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O‑Glycopeptides from Whole Cell Proteomess. J. Proteome Res., 16: 1706−18. [PubMed]
  2. Woo, C.M., et al. (2017). Mapping and Quantification of Over 2000 O-linked Glycopeptides in Activated Human T Cells with Isotope-Targeted Glycoproteomics (Isotag). Mol. Cell. Proteomics 17: 764-75. [PubMed]
  3. Goa, G., et al. (2017). Small Molecule Interactome Mapping by Photoaffinity Labeling Reveals Binding Site Hotspots for the NSAIDs. J. Am. Chem. Soc., 140: 4259−68. [PubMed]
  4. Woo, C.M., et al. (2015). Isotope-targeted glycoproteomics (IsoTaG): a mass-independent platform for intact N- and O-glycopeptide discovery and analysis.Nat Methods., 12: 561-7. [PubMed]
  5. Weerapana, E., et al. (2010). Quantitative reactivity profiling predicts functional cysteines in proteomes. Nature 468: 790−5. [PubMed]

IsoStamp v2

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]]> Ulex Europaeus Agglutinin I (UEA I), DyLight® 649 https://staging.vectorlabs.com/products/dylight-649-ulex-europaeus-agglutinin/ Sat, 06 May 2023 01:31:23 +0000 https://staging.vectorlabs.com/?post_type=product&p=15240 Description

Ulex europaeus agglutinin I binds to many glycoproteins and glycolipids containing α-linked fucose residues, such as ABO blood group glycoconjugates. This lectin preferentially binds blood group O cells and has been used to determine secretor status. It has been established as an excellent marker for human endothelial cells.

DyLight 594 labeled Ulex europaeus agglutinin I has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation at 655 nm
  • Emission at 670 nm
  • Color: Far Red

Specifications

Unit Size 1 mg
Applications Immunofluorescence, Glycobiology
Recommended Usage The recommended concentration range for use is 5-20 µg/ml. If a precipitate forms upon long-term storage, warm to 37 ºC.
Recommended Storage 2-8 °C
Maximum Excitation 655 nm
Maximum Emission 670 nm
Solution 10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2.
Concentration 1 mg active conjugate/ml
Conjugate DyLight 649
Color of Fluorescence Far Red
Sugar Specificity Fucose, Arabinose

The post Ulex Europaeus Agglutinin I (UEA I), DyLight® 649 appeared first on VectorLabs.

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Description

Ulex europaeus agglutinin I binds to many glycoproteins and glycolipids containing α-linked fucose residues, such as ABO blood group glycoconjugates. This lectin preferentially binds blood group O cells and has been used to determine secretor status. It has been established as an excellent marker for human endothelial cells.

DyLight 594 labeled Ulex europaeus agglutinin I has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation at 655 nm
  • Emission at 670 nm
  • Color: Far Red

Specifications

Unit Size1 mg
ApplicationsImmunofluorescence, Glycobiology
Recommended UsageThe recommended concentration range for use is 5-20 µg/ml. If a precipitate forms upon long-term storage, warm to 37 ºC.
Recommended Storage2-8 °C
Maximum Excitation655 nm
Maximum Emission670 nm
Solution10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2.
Concentration1 mg active conjugate/ml
ConjugateDyLight 649
Color of FluorescenceFar Red
Sugar SpecificityFucose, Arabinose

Documents

Citations

Technical Information

Accompanying each fluorescent lectin is an analysis data sheet summarizing the results of our quality control tests and providing pertinent information on the product. All of these reagents are supplied as solutions preserved with sodium azide.

Inhibiting/Eluting Sugar: 50 mM – 100 mM L-fucose

The post Ulex Europaeus Agglutinin I (UEA I), DyLight® 649 appeared first on VectorLabs.

]]> Sambucus Nigra Lectin (SNA, EBL), CY5 https://staging.vectorlabs.com/products/cy5-sambucus-nigra-lectin/ Tue, 02 May 2023 19:59:19 +0000 https://staging.vectorlabs.com/?post_type=product&p=14594 Description

Sambucus nigra lectin, isolated from elderberry bark, binds preferentially to sialic acid attached to terminal galactose in α-2,6 and to a lesser degree, α-2,3 linkage. Binding is also inhibited to some extent by lactose or galactose. This lectin appears to bind sialic acid linked to N-acetylgalactosamine or galactose. SNA has been reported to inhibit cell-free protein synthesis.

Cy5 labeled Sambucus nigra lectin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation:  650 nm
  • Emission:  670 nm
  • Color:  Far Red

Specifications

Unit Size 1 mg
Applications Immunofluorescence, Glycobiology
Recommended Usage The recommended concentration range for use is 5-20 µg/ml.
Recommended Storage 2-8 °C
Maximum Excitation 650 nm
Maximum Emission 670 nm
Solution 10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer
Concentration 1 mg active conjugate/ml
Conjugate Cy5
Color of Fluorescence Far Red
Sugar Specificity Sialic Acid

The post Sambucus Nigra Lectin (SNA, EBL), CY5 appeared first on VectorLabs.

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Description

Sambucus nigra lectin, isolated from elderberry bark, binds preferentially to sialic acid attached to terminal galactose in α-2,6 and to a lesser degree, α-2,3 linkage. Binding is also inhibited to some extent by lactose or galactose. This lectin appears to bind sialic acid linked to N-acetylgalactosamine or galactose. SNA has been reported to inhibit cell-free protein synthesis.

Cy5 labeled Sambucus nigra lectin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes. 

  • Excitation:  650 nm
  • Emission:  670 nm
  • Color:  Far Red
 

Specifications

Unit Size1 mg
ApplicationsImmunofluorescence, Glycobiology
Recommended UsageThe recommended concentration range for use is 5-20 µg/ml.
Recommended Storage2-8 °C
Maximum Excitation650 nm
Maximum Emission670 nm
Solution10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer
Concentration1 mg active conjugate/ml
ConjugateCy5
Color of FluorescenceFar Red
Sugar SpecificitySialic Acid

Documents

Technical Information

Accompanying each fluorescent lectin is an analysis data sheet summarizing the results of our quality control tests and providing pertinent information on the product. All of these reagents are supplied as solutions preserved with sodium azide. 
Elution: 500 mM lactose in buffered saline followed by 500 mM lactose in acetic acid

The post Sambucus Nigra Lectin (SNA, EBL), CY5 appeared first on VectorLabs.

]]> Sambucus Nigra Lectin (SNA, EBL), CY3 https://staging.vectorlabs.com/products/cy3-labeled-sambucus-nigra-lectin/ Tue, 02 May 2023 19:52:53 +0000 https://staging.vectorlabs.com/?post_type=product&p=14583 Description

Sambucus nigra lectin, isolated from elderberry bark, binds preferentially to sialic acid attached to terminal galactose in α-2,6 and to a lesser degree, α-2,3 linkage. Binding is also inhibited to some extent by lactose or galactose. This lectin appears to bind sialic acid linked to N-acetylgalactosamine or galactose. SNA has been reported to inhibit cell-free protein synthesis.

Cy3 labeled Sambucus nigra lectin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation:  552 nm
  • Emission:  565 nm
  • Color:  Red

Specifications

Unit Size 1 mg
Applications Immunofluorescence, Glycobiology
Recommended Usage The recommended concentration range for use is 5-20 µg/ml.
Recommended Storage 2-8 °C
Maximum Excitation 552 nm
Maximum Emission 565 nm
Solution 10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer.
Concentration 1 mg active conjugate/ml
Conjugate Cy3
Color of Fluorescence Red
Sugar Specificity Sialic Acid

The post Sambucus Nigra Lectin (SNA, EBL), CY3 appeared first on VectorLabs.

]]>

Description

Sambucus nigra lectin, isolated from elderberry bark, binds preferentially to sialic acid attached to terminal galactose in α-2,6 and to a lesser degree, α-2,3 linkage. Binding is also inhibited to some extent by lactose or galactose. This lectin appears to bind sialic acid linked to N-acetylgalactosamine or galactose. SNA has been reported to inhibit cell-free protein synthesis.

Cy3 labeled Sambucus nigra lectin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes. 

  • Excitation:  552 nm
  • Emission:  565 nm
  • Color:  Red

Specifications

Unit Size1 mg
ApplicationsImmunofluorescence, Glycobiology
Recommended UsageThe recommended concentration range for use is 5-20 µg/ml.
Recommended Storage2-8 °C
Maximum Excitation552 nm
Maximum Emission565 nm
Solution10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer.
Concentration1 mg active conjugate/ml
ConjugateCy3
Color of FluorescenceRed
Sugar SpecificitySialic Acid

Documents

Technical Information

Accompanying each fluorescent lectin is an analysis data sheet summarizing the results of our quality control tests and providing pertinent information on the product. All of these reagents are supplied as solutions preserved with sodium azide. 
Elution: 500 mM lactose in buffered saline followed by 500 mM lactose in acetic acid

The post Sambucus Nigra Lectin (SNA, EBL), CY3 appeared first on VectorLabs.

]]> Peanut Agglutinin (PNA), CY5 https://staging.vectorlabs.com/products/cy5-peanut-agglutinin/ Tue, 02 May 2023 19:49:30 +0000 https://staging.vectorlabs.com/?post_type=product&p=14573 Description

Peanut agglutinin binds preferentially to the T-antigen, a galactosyl (β-1,3) N-acetylgalactosamine structure present in many glycoconjugates such as M and N blood groups, gangliosides, and many other soluble and membrane-associated glycoproteins and glycolipids. With certain exceptions, the receptor sequence for PNA is normally sialylated which prevents the lectin from binding to its receptor oligosaccharide (see Jacalin). Even sialic acid which is not bound directly to the receptor sugars may inhibit binding. The presence of calcium ions in diluents can enhance the binding of PNA to receptors, possibly by neutralizing the negative charges on sialic acid residues adjacent to the receptor sequence.

Cy5 labeled Peanut agglutinin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation:  650 nm
  • Emission:  670 nm
  • Color:  Far Red

Specifications

Unit Size 1 mg
Applications Immunofluorescence, Glycobiology
Recommended Usage The recommended concentration range for use is 5-20 µg/ml.
Recommended Storage 2-8 °C
Maximum Excitation 650 nm
Inhibiting and/or Eluting Sugar Inhibiting/Eluting Sugar: 200 mM galactose (S-9003).
Maximum Emission 670 nm
Solution 10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer.
Concentration 1 mg active conjugate/ml
Conjugate Cy5
Color of Fluorescence Far Red
Sugar Specificity Galactose

 

The post Peanut Agglutinin (PNA), CY5 appeared first on VectorLabs.

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Description

Peanut agglutinin binds preferentially to the T-antigen, a galactosyl (β-1,3) N-acetylgalactosamine structure present in many glycoconjugates such as M and N blood groups, gangliosides, and many other soluble and membrane-associated glycoproteins and glycolipids. With certain exceptions, the receptor sequence for PNA is normally sialylated which prevents the lectin from binding to its receptor oligosaccharide (see Jacalin). Even sialic acid which is not bound directly to the receptor sugars may inhibit binding. The presence of calcium ions in diluents can enhance the binding of PNA to receptors, possibly by neutralizing the negative charges on sialic acid residues adjacent to the receptor sequence.

Cy5 labeled Peanut agglutinin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation:  650 nm
  • Emission:  670 nm
  • Color:  Far Red

Specifications

Unit Size1 mg
ApplicationsImmunofluorescence, Glycobiology
Recommended UsageThe recommended concentration range for use is 5-20 µg/ml.
Recommended Storage2-8 °C
Maximum Excitation650 nm
Inhibiting and/or Eluting SugarInhibiting/Eluting Sugar: 200 mM galactose (S-9003).
Maximum Emission670 nm
Solution10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer.
Concentration1 mg active conjugate/ml
ConjugateCy5
Color of FluorescenceFar Red
Sugar SpecificityGalactose

Documents

Technical Information

 

PNA is useful in distinguishing between normal and tumor tissues and in assessing malignancy in transitional mucosa. In addition, PNA binding can be used to measure cellular maturity in lymphoid tissues, to distinguish a variety of lymphocyte subpopulations in man and experimental animals, and to measure the levels of lymphoid cell populations in many diseases. PNA can be employed in the fractionation of stem cells in mice for use in bone marrow transplantation across histocompatibility barriers.

A major cell surface receptor for PNA may be asialo GM1 ganglioside. Since PNA shares specificity with the antibody to this glycolipid, PNA and the antibody can be used interchangeably in some applications.

Accompanying each fluorescent lectin is an analysis data sheet summarizing the results of our quality control tests and providing pertinent information on the product. All of these reagents are supplied as solutions preserved with sodium azide.

Inhibiting/Eluting Sugar: 200 mM galactose

The post Peanut Agglutinin (PNA), CY5 appeared first on VectorLabs.

]]> Peanut Agglutinin (PNA), CY3 https://staging.vectorlabs.com/products/cy3-peanut-agglutinin/ Tue, 02 May 2023 19:46:03 +0000 https://staging.vectorlabs.com/?post_type=product&p=14562

Description

Peanut agglutinin binds preferentially to the T-antigen, a galactosyl (β-1,3) N-acetylgalactosamine structure present in many glycoconjugates such as M and N blood groups, gangliosides, and many other soluble and membrane-associated glycoproteins and glycolipids. With certain exceptions, the receptor sequence for PNA is normally sialylated which prevents the lectin from binding to its receptor oligosaccharide (see Jacalin). Even sialic acid which is not bound directly to the receptor sugars may inhibit binding. The presence of calcium ions in diluents can enhance the binding of PNA to receptors, possibly by neutralizing the negative charges on sialic acid residues adjacent to the receptor sequence.

Cy3 labeled Peanut agglutinin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation: 552 nm
  • Emission: 565 nm
  • Color: Red

Specifications

Unit Size 1 mg
Applications Immunofluorescence, Glycobiology
Recommended Usage The recommended concentration range for use is 5-20 µg/ml.
Recommended Storage 2-8 °C
Maximum Excitation 552 nm
Inhibiting and/or Eluting Sugar Inhibiting/Eluting Sugar: 200 mM galactose (S-9003).
Maximum Emission 565 nm
Solution 10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer.
Concentration 1 mg active conjugate/ml
Conjugate Cy3
Color of Fluorescence Red
Sugar Specificity Galactose

The post Peanut Agglutinin (PNA), CY3 appeared first on VectorLabs.

]]>

Description

Peanut agglutinin binds preferentially to the T-antigen, a galactosyl (β-1,3) N-acetylgalactosamine structure present in many glycoconjugates such as M and N blood groups, gangliosides, and many other soluble and membrane-associated glycoproteins and glycolipids. With certain exceptions, the receptor sequence for PNA is normally sialylated which prevents the lectin from binding to its receptor oligosaccharide (see Jacalin). Even sialic acid which is not bound directly to the receptor sugars may inhibit binding. The presence of calcium ions in diluents can enhance the binding of PNA to receptors, possibly by neutralizing the negative charges on sialic acid residues adjacent to the receptor sequence.

Cy3 labeled Peanut agglutinin has an appropriate number of fluorochromes bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated fluorochromes.

  • Excitation: 552 nm
  • Emission: 565 nm
  • Color: Red

Specifications

Unit Size1 mg
ApplicationsImmunofluorescence, Glycobiology
Recommended UsageThe recommended concentration range for use is 5-20 µg/ml.
Recommended Storage2-8 °C
Maximum Excitation552 nm
Inhibiting and/or Eluting SugarInhibiting/Eluting Sugar: 200 mM galactose (S-9003).
Maximum Emission565 nm
Solution10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2, and a proprietary stabilizer.
Concentration1 mg active conjugate/ml
ConjugateCy3
Color of FluorescenceRed
Sugar SpecificityGalactose

Documents

Technical Information

PNA is useful in distinguishing between normal and tumor tissues and in assessing malignancy in transitional mucosa. In addition, PNA binding can be used to measure cellular maturity in lymphoid tissues, to distinguish a variety of lymphocyte subpopulations in man and experimental animals, and to measure the levels of lymphoid cell populations in many diseases. PNA can be employed in the fractionation of stem cells in mice for use in bone marrow transplantation across histocompatibility barriers.

A major cell surface receptor for PNA may be asialo GM1 ganglioside. Since PNA shares specificity with the antibody to this glycolipid, PNA and the antibody can be used interchangeably in some applications.

Accompanying each fluorescent lectin is an analysis data sheet summarizing the results of our quality control tests and providing pertinent information on the product. All of these reagents are supplied as solutions preserved with sodium azide.

Inhibiting/Eluting Sugar: 200 mM galactose

The post Peanut Agglutinin (PNA), CY3 appeared first on VectorLabs.

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