Biotin – VectorLabs

Streptavidin Agarose Ultra Performance™

Vector Laboratories R&D — Wed, 03 May 2023 20:45:49 +0000

Description

Streptavidin Agarose Ultra Performance provides high biotin binding capacity at a low price. We used our ChromaLINK bioconjugation technology to crosslink high activity streptavidin to uniform 35 micron, 6% highly crosslinked agarose. This ideal combination provides a biotin binding capacity of >330 nmol/mL of resin—the highest binding capacity in the industry. The base agarose beads support high flow rates and backpressures, which is ideal for automation in packed cartridges. Use Streptavidin Agarose Ultra Performance for improved recovery of any biotinylated biomolecule with lower non-specific binding.

Specifications

Bead Size	35 µm
Loading Capacity	≥ 330 nmol/mL
Reactivity	Biotin
Applications	Antibody Labeling, Aptamers, Next-Generation Sequencing (NGS), Photocrosslinking Studies
Recommended Storage	2° – 8°C – Do Not Freeze

Documents

Citations

Technical Information

The Streptavidin Agarose Ultra Performance is a high biotin binding-capacity resin used for the immobilization of biotinylated biomolecules. High specific-activity recombinant streptavidin is immobilized on 6% highly-crosslinked beaded agarose using the SoluLINK proprietary linking technology. With a biotin binding capacity of >330nmol / mL resin, Streptavidin Agarose is one of the highest loading capacity resins currently available.

Legal Information

SoluLINK^® Bioconjugation For Research Use Only. Not for use in diagnostic procedures. For additional licensing restrictions, please see the license agreement at vectorlabs.com/solulink-research-license.

Products are for research use only, not for use in diagnostic or therapeutic procedures or for use in humans. Products are not for resale without express written permission of Seller. No license under any patent or other intellectual property right of Seller or its licensors is granted or implied by the purchase unless otherwise provided in writing.

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MagnaLINK® Streptavidin Magnetic Beads (2.8 µm)

Vector Laboratories R&D — Wed, 03 May 2023 20:40:00 +0000

Description

MagnaLINK 2.8 µm Streptavidin Magnetic Beads offer the highest biotin binding capacity on the market, combined with exceptional size uniformity. Higher binding translates to reduced bead mass required to immobilize a biotinylated sample and lower background from nonspecific binding, resulting in better signals and lower costs. MagnaLINK Streptavidin Magnetic Beads are 2.8 micron diameter, super-paramagnetic, hydrophilic polymer–encapsulated (no exposed iron), monodispersed microspheres with a uniform size distribution and a fast (<2 min) magnetic response time. They are colloidally stable with or without non-ionic detergents. The key to high biotin binding is in the unique covalently crosslinked streptavidin, based on ChromaLINK^® technology. The high surface area, when combined with our efficient linking chemistry, produces a consistent product with a free biotin binding capacity of at least 12 nmol per milligram of beads. MagnaLINK beads also demonstrate exceptional size uniformity of <5% CV, as evidenced by scanning electron microscopy (SEM). These combined technologies make MagnaLINK beads the ideal magnetic bead for high-throughput robotic applications.

Specifications


Bead Size	2.8 µm
Loading Capacity	≥ 10 nmol/mg
Reactivity	Biotin
Applications	Antibody Labeling, Aptamers, Next-Generation Sequencing (NGS), Photocrosslinking Studies
Recommended Storage	2° – 8°C – Do Not Freeze

Documents

Citations

Technical Information

Product Description

MagnaLINK Streptavidin Magnetic beads are highly uniform, 2.8 µmeter polymer-encapsulated (no exposed iron), super-paramagnetic beads containing covalently cross-linked streptavidin (Figure 1). MagnaLINK streptavidin magnetic beads are made by covalently cross-linking streptavidin to a hydrophilic surface using the SoluLINK conjugation technology. The high surface area of these paramagnetic beads when combined with the efficient SoluLINK linking technology produces the most consistent and highest biotin binding capacity (≥10 nmol/mg) of any uniform streptavidin magnetic bead on the market. Beads are supplied at 1% solids (10 mg/mL) in nuclease-free water with 0.05% sodium azide.

Figure 1. Scanning electron microscope image of MagnaLINK Streptavidin Magnetic beads.

Figure 2. Cross-sectional diagram of MagnaLINK Streptavidin Magnetic bead.

Features of our MagnaLINK beads

Highest free biotin binding capacity of any uniform bead (≥ 10 nmol/mg)
Binds 0.8 nmol/mg biotinylated oligonucleotide
Binds 0.75 nmol/mg biotinylated-IgG @ 4 biotins/IgG
Beads are encapsulated (no exposed iron)
Paramagnetic beads are highly uniform in size (2.8 ± 0.2 microns)
Fast magnetic response time (60% w/w magnetite)

Binding Capacity Comparison: MagnaLINK Streptavidin Magnetic Bead Vs. Competitor’s Bead of Similar Size

Ligand	MagnaLINK 2.8 µm Binding Capacity	Competitor’s 2.8 µm Bead Binding Capacity
Free biotin	≥ 10 nmol/mg	0.90 nmol/mg
Biotinylated Oligo (23-mer)	≥ 0.8 nmol/mg	N/A
Biotinylated IgG (4 biotins per IgG)	750 pmol/mg (113 µg/mg)	60 pmol/mg (10 µg/mg)

MagnaLINK References:

Moore R, Worrallo M, Mitchell P, Harriman J, Glen K, Thomas R. Immobilisation of Delta-like 1 ligand for the scalable and controlled manufacture of hematopoietic progenitor cells in a stirred bioreactor. BMC Biotechnology volume 17, Article number: 65 (2017).

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NanoLINK® Streptavidin Magnetic Beads (1.0 µm)

Vector Laboratories R&D — Wed, 03 May 2023 20:26:26 +0000

Description

NanoLINK 1.0 µm Streptavidin Magnetic Beads have the highest biotin binding capacity on the market. Higher binding translates into reduced bead mass required to immobilize a biotinylated sample and lower background noise from nonspecific binding, resulting in better signals and lower net costs. NanoLINK Streptavidin Magnetic Beads are 1.0 µm in diameter and bind at least 12 nmol of free biotin per milligram of beads.

Specifications

Bead Size	1 μm
Loading Capacity	≥ 12 nmol/mg
Reactivity	Biotin
Applications	Antibody Labeling, Aptamers, Next-Generation Sequencing (NGS), Photocrosslinking Studies
Recommended Storage	2° – 8°C – Do Not Freeze

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Citations

Technical Information

Product Description

NanoLINK Streptavidin Magnetic Beads are nanometer-sized, polymer-encapsulated (no exposed iron), super-paramagnetic particles containing covalently cross-linked streptavidin. NanoLINK streptavidin magnetic beads are made by covalently cross-linking streptavidin to a hydrophilic surface using the SoluLINK conjugation technology. The high surface area of these magnetic beads when combined with the efficient SoluLINK linking technology produces the most consistent and highest biotin binding capacity per unit mass of any streptavidin magnetic bead on the market.

Cross-sectional diagram of NanoLINK Streptavidin Magnetic bead.

Product Features

Highest free biotin binding capacity of any commercially available bead (≥ 12 nmol/mg)
Binds ≥ 2.5 nmol/mg of a biotinylated oligonucleotide
Binds ≥ 1.7 nmol/mg of a biotinylated-IgG (250 μg/mg) @ 3 biotins/IgG
Encapsulated (no exposed iron) beads
Bead size is less than 875 nm
Fast magnetic response time (25 electromagnetic unit)
Paramagnetic (no residual magnetism)

Binding Capacity Comparison: NanoLINK Streptavidin Magnetic Bead Vs. Competitor’s Bead of Similar Size

Ligand	NanoLINK 0.8 µm Binding Capacity	Competitor’s 1.0 µm Bead Binding Capacity
Free biotin	≥ 12 nmol/mg	1.3 nmol/mg
Biotinylated Oligo (23-mer)	≥ 2.5 nmol/mg	N/A
Biotinylated IgG (4 biotins per IgG)	≥ 1.7 nmol/mg (250 µg/mg)	0.07 nmol/mg (10 µg/mg)

Applications

NanoLINK streptavidin magnetic beads possess the highest biotin-binding capacity of any commercially available polymer-encapsulated streptavidin bead. These beads are particularly suited for high throughput robotic applications where high biotin loads must be immobilized and separated using a suitably strong magnet.

The beads can be used to immobilize:

Biotinylated antibodies and other proteins
Biotinylated dsDNA (gDNA, PCR products) or biotinylated aRNA
Biotinylated oligonucleotides

The main advantage of using these ultra-high capacity beads is that it leads to a reduction in the overall particle mass required to immobilize a biotinylated sample. This in turn leads to reduced costs and lower non-specific background (NSB).

Applications include separation of biotin-labeled biomolecules such as biotin- labeled antibodies, genomic DNA, RNA, PCR products, oligonucleotides (e.g. biotinylated oligo (dT) or peptides. NanoLINK streptavidin magnetic beads are also ideal for generating single-stranded PCR templates (by removal of the unbiotinylated competing PCR strand), which allows for a dramatic increase in the efficiency of hybridization to complementary targets.

NanoLINK streptavidin magnetic beads also provide an affordable alternative for automated, high throughput immobilization processes that utilize 96-well magnets to affect multiplex binding and separation of nucleic acid or immunoassay biomolecules. NanoLINK beads are supplied at 10 mg/mL, with 1% solids, in nuclease-free water containing 0.05% sodium azide. There are no surfactants present although particles can be washed with water prior use to remove residual azide if desired.

Bead Characteristics

Percent Solids: NanoLINK streptavidin magnetic beads are packaged nominally at 1% solids (10 mg/mL) as measured using spectroscopic analysis set by their optical density at 600 nm versus a known mass standard of the same size.

Biotin Binding Capacity: The biotin binding capacity of NanoLINK streptavidin magnetic beads is measured in nmol/mg. Biotin binding is quantitatively measured by incubating a known mass of beads (0.5 mg) with a fluorescein-biotin standard solution for 60 minutes and quantifying the amount of residual unbound fluorescein-biotin left in solution versus negative control beads.

Size Distribution by Scanning Electron Microscopy (SEM): Scanning electron microscopy confirms a bimodal size distribution for NanoLINK Magnetic Beads. The streptavidin beads consist of a core polymeric particle ~ 850 nm surrounded by a population of similar but smaller polymeric beads ~ 150 nm.

Quality control: A sample of each NanoLINK batch is retained for biotin binding capacity and % solids analysis. The remainder is packaged, refrigerated, and preserved in nuclease-free water containing 0.05% sodium azide to prevent microbial contamination.

Cleaning: Surfactants are not added to this product and the particles are thoroughly washed with nuclease-free water containing 0.05% sodium azide prior to packaging. For some applications it may be desirable to remove residual azide using a brief wash.

Stability: Particles should be stored at 2-8^o C. Do not freeze. If beads are settled, resuspend by suitable methods including: vortexing, rotary mixing, or swirling. NanoLINK streptavidin magnetic beads remain stable when stored at 2-8^o C for 1 year.

Washing: NanoLINK streptavidin magnetic beads are washed by magnetic separation using commercially available magnetic stands. Magnetic stands are available in 50 mL, 15 mL, 1.5 mL, and 96-well plate formats from various vendors. NanoLINK beads are placed on a magnetic stand for 2-3 minutes and the clarified supernatant removed without disturbing the pellet.

Re-suspension: After long-term storage and settling of beads, it is best to resuspend the beads thoroughly to avoid any mild bead-to-bead aggregation.

NanoLINK References:

Viktorovskaya O, Greco T, Cristea I, and S. Thompson. Identification of RNA Binding Proteins Associated with Dengue Virus RNA in Infected Cells Reveals Temporally Distinct Host Factor Requirements. PLoS Neglected Tropical Diseases 2016 Aug; 10(8): e0004921.
Beaumont M, Tomazela D, and E. Escandón. Antibody-drug conjugates: integrated bioanalytical and biodisposition assessments in lead optimization and selection. AAPS Open volume 4, Article number: 6 (2018).
Zhao Y, Valbuena G, Walker D, Gazi M, Hidalgo M, DeSousa R, Antonio J, Oteo Y.G., and A.R. Brasier. Endothelial Cell Proteomic Response to Rickettsia conorii Infection Reveals Activation of the Janus Kinase (JAK)-Signal Transducer and Activator of Transcription (STAT)-Inferferon Stimulated Gene (ISG)15 Pathway and Reprogramming Plasma Membrane Integrin/Cadherin Signaling. Molecular & Cellular Proteomics. 15 (1) 289-304, 2016.

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ChromaLINK® Biotin Protein Labeling

Vector Laboratories R&D — Wed, 03 May 2023 04:40:32 +0000

Description

The ChromaLINK Biotin Protein Labeling Kit includes everything you need to biotinylate your protein: buffers, reagents, spin columns, and an easy-to-follow protocol and online calculator. With the ChromaLINK Biotin Protein Labeling Kit you can biotinylate and purify up to 5 antibodies or other proteins ranging in molecular weight from 20–200 kDa and from as little as 25 µg to as much as 1 mg.

Specifications

Conjugation Targets	Antibody
Label/Modifier Type	biotin
Reactivity	Amine
Recommended Storage	2° – 8°C — Do Not Freeze
Applications	Antibody Labeling

Kit Components

ChromaLINK Biotin (5 x 0.5 mg)
10X Modification Buffer (2 x 1.5 mL)
10X PBS (2 x 1.5 mL)
0.5 mL Zeba™ Column (12)
2 mL Collection Tube (24)
1.5 mL Storage Tube (12)
Bovine IgG Control (0.5 mg)
Biotinylated Bovine IgG Control (0.5 mg)
Anhydrous DMF (1.5 mL)

Documents

Technical Information

A. Product Description

The ChromaLINK Biotin Labeling kit has all the necessary reagents for the traceable biotinylation of any lysine-containing protein. This kit provides sufficient materials to biotinylate and purify up to 5 proteins in about 2 hours. The scale of each protein labeling reaction is chosen by the researcher and can span from as little as 25 μg to as much as 1 milligram of protein per reaction.

The ChromaLINK Biotin labeling process is based on a patented UV-traceable linker called ChromaLINK Biotin (Figure 1). This process is based on an advanced protein biotinylation reagent containing an aromatic, N-hydroxy-succinimidyl ester functional group (a), which efficiently modifies protein lysine residues in a phosphate buffer system. The linker possesses an embedded bis-aryl hydrazone structure (b), which forms the compound’s UV-traceable chromophore. The traceable nature of this unique chromophore enables the non-destructive quantification of biotin (c), attached to the protein. These three diverse chemical elements are linked together through a long-chain PEG3 spacer (d), which preserves streptavidin/biotin affinity while simultaneously maintaining antibody solubility.

Figure 1. Structure of ChromaLINK Biotin C38H50N8O10S; MW: 810.92

b. Benefits and Features

The ChromaLINK Biotin Labeling kit offers an easier solution to the quantitative biotinylation of proteins starting from as little as 25 μg to as much as 1 milligram. Proteins ranging in molecular weights from 20–200 kDa are readily biotinylated using ChromaLINK Biotin. The accompanying instructions and protocols are designed to make traceable biotinylation of proteins easy to perform. The use of ChromaLINK Biotin eliminates the need to carry out cumbersome and indirect biotin binding assays such as the time consuming HABA or FluoroReporter™ assay used to determine the degree of biotin incorporation. Rather, after labeling, the biotin molar substitution ratio is measured directly by a simple UV-spectrophotometric scan (220–420 nm) or single point measurements at two wavelengths (A280/A354). Alternatively, a single wavelength (A354) can be used in combination with a protein assay (e.g., BCA or Bradford) to quantify the molar substitution ratio (MSR) or number of biotins per protein molecule.

c. Protocol Summary

Sample preparation (10 min)
1st buffer exchange (10 min)
Protein determination assay (BCA assay) (30 min)
Biotinylation (120 min)
2nd buffer exchange (10 min)
UV analysis and calculations (20 min)

d. Labeling Process Diagram

The ChromaLink Biotin protein labeling process is diagramed in Figure 2.

Figure 2. ChromaLink Biotin protein labeling process.

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ChromaLINK® One-Shot™ Antibody Biotinylation Kit

Vector Laboratories R&D — Wed, 03 May 2023 04:32:38 +0000

Description

The ChromaLINK One-Shot Antibody Biotinylation Kit provides convenient, consistent, and measurable biotinylation of 100 µg of antibody. Each kit contains ChromaLINK Biotin, which incorporates a novel UV-traceable chromophore in the linker arm to enable reproducibility in your biotinylation process. Now you can measure the degree of biotinylation in minutes, not hours, without the standard curves required for the HABA/Avidin and Fluoro-Reporter assays. The ChromaLINK One-Shot kit contains everything needed to to biotinylate your antibody: buffers, reagents, desalting columns, and an easy-to-follow protocol and online biotin incorporation calculator.

Specifications

Conjugation Targets	Antibody
Label/Modifier Type	biotin
Reactivity	Amine
Recommended Storage	2° – 8°C — Do Not Freeze
Applications	Antibody Labeling, In Situ Proximity Ligation
Unit Size	1 Kit

Kit Components

ChromaLINK Biotin (6.49 μg)
1X Modification Buffer (1.5 mL)
1X PBS (2 x 1.5 mL)
2 mL Collection Tube (4)
0.5 mL Zeba™ Desalting Column (2)
Biotinylated Bovine IgG Control (100 μg)
1M Tris HCI (1.5 mL)
Anhydrous DMF (1.5 mL)

Documents

Product FAQs

For B-9007-105K, why you would need to perform buffer exchange on the protein if you have reconstituted it in the 1x modification buffer?

The purpose of the buffer exchange step is to remove amine contaminants, such as Tris or glycine, from the protein. These contaminants may interfere with NHS ester labeling, which targets primary amine groups. Some samples are so excessively contaminated that two buffer exchanges may be required before proceeding with biotinylation.

Citations

Technical Information

A. Product Description

The ChromaLINK Biotin One-Shot Antibody Labeling Kit is designed to biotinylate a single 100 microgram quantity of antibody in about 2.0 hours. The ChromaLINK One-Shot kit relies on a UV-traceable linker called ChromaLINK Biotin to label the antibody (Figure 1). This unique labeling reagent contains an aromatic, water-soluble N-hydroxy-succinimidyl ester functional group. (a) that efficiently modifies antibody lysine residues under mild phosphate buffer conditions. The reagent also possesses an embedded bis-aryl hydrazone structure (b) which forms the linker’s UV-traceable chromophore. The absorbance signature provided by the chromophore enables rapid and non-destructive quantification of incorporated biotin. (c). The linker also features a long PEG3 spacer (d) which preserves streptavidin/biotin affinity and helps maintain antibody solubility.

Figure 1. Structure of ChromaLINK Biotin C38H50N8O10S; MW 810.92.

B. Features and Benefits

The ChromaLINK Biotin One-Shot Kit is a simple, cost-effective way of incorporating a readily verifiable amount of biotin into a single 100 μg quantity of antibody. The kit conveniently features a pre-weighed amount of ChromaLINK Biotin reagent that is readily resuspended directly into the antibody solution. ChromaLINK Biotin can be used to label a variety of different antibodies, including mammalian IgG (monoclonal or polyclonal) and/or avian IgY. Biotin incorporation is rapidly determined by means of a simple, non-destructive UV-scan (220–400 nm) of the sample after removal of excess labeling reagent. The kit features high antibody recovery (60–90 μg) and a consistent level of biotin incorporation (3–8 biotin molecules per antibody molecule) when used as directed.

C. ChromaLINK One-Shot Process Diagram

The ChromaLINK Biotin One-Shot antibody labeling procedure is illustrated in Figure 2.

Figure 2. ChromaLINK Biotin One-Shot antibody labeling procedure.

D. Process Summary

Sample Preparation: bring antibody to 1 mg/mL in 100 μL buffer
1st Sample Analysis: confirm antibody concentration using spectrophotometer
1st Buffer Exchange: equilibrate spin columns and buffer exchange antibody
2nd Sample Analysis: reconfirm antibody concentration using spectrophotometer
Biotinylation: label antibody with ChromaLINK Biotin
2nd Buffer Exchange: remove excess labeling reagent
Quantify Biotin Incorporation: measure biotin incorporation using spectrophotometer

E. Important Labeling Parameters

The ChromaLINK Biotin One-Shot Antibody Labeling Kit is designed to biotinylate a single 100 μg quantity of antibody resuspended in 100 μL buffer as indicated in Table 1.

Table 1. Initial conditions required for the ChromaLINK Biotin One-Shot labeling procedure.
This kit provides a consistent and reliable degree of biotinylation by controlling the following reaction variables:

Fixed antibody mass (100 μg) and volume (100 μL)
Optimized reaction buffer
Controlled and fixed reaction time (90 min)
Consistent reaction stoichiometry (12X mole-equivalents) (12-fold molar excess of biotin reagent)

Critical to consistent biotin incorporation is the ability of a user to accurately determine the antibody’s initial protein concentration in a non-destructive manner that allows full or nearly full recovery of the precious sample. The ChromaLINK One-Shot procedure recommends using a scan (220–400 nm) on a spectrophotometer rather than the traditional A280 to estimate antibody concentration. The rationale for using a scan rather than a single wavelength (A280) is that many commercial antibodies contain preservatives and/or other additives that mask or distort the intrinsic A280 of a sample. The presence of an additive can make it more difficult or sometimes impossible to accurately estimate protein concentration via A280. A scan however contains greater spectral information, often revealing the presence of spectrum-altering additives that are known to interfere with the accurate estimate of protein concentration. Altered spectra are a warning that the A280 of the sample will not yield an accurate estimate of protein concentration. Commonly used additives include preservatives such as sodium azide, thimerosal, protein stabilizers such as BSA or gelatin and/or small molecule additives such as glycine or trehalose.

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ChromaLINK® Biotin Maleimide

Vector Laboratories R&D — Wed, 03 May 2023 04:21:45 +0000

Description
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Description

ChromaLINK Biotin contains a UV-traceable chromophore based on ChromaLINK technology to enable reproducibility in your biotinylation process. Now you can measure the degree of biotinylation in minutes, not hours, without the standard curves required for HABA/avidin and fluoro-reporter assays. With a simple UV scan, you can quantify biotin incorporation and ensure reproducible production of consistent batches. The maleimide group reacts with free thiol groups on biomolecules and surfaces, such as the hinge region of antibodies after mild reduction of sulfhydryl groups with TCEP or DTT.

Specifications

Label/Modifier Type	biotin
Reactivity	Streptavidin
Recommended Storage	Desiccated: -15° to -25°C
Applications	Antibody Labeling, Aptamers
Other Name(s)	SoluLINK Bioconjugation

Documents

Technical Information

Introduction to ChromaLINK Labeling Technology

ChromaLINK Biotin Maleimide incorporates UV-traceable biotin onto thiol containing proteins, peptides and/or antibodies. ChromaLINK Biotin Maleimide has been engineered to include many novel features. As illustrated in Figure 1, the molecule’s structure contains a bis-aryl hydrazone chromophore (a), linked by a PEG3 linker arm (b), to biotin (c). This reagent permits direct spectroscopic quantification of incorporated biotin. The extended PEG3 linker preserves biotin/streptavidin affinity and maintains protein solubility after modification while the maleimide functional group (d), efficiently modifies thiols in aqueous buffers.

Figure 1. Molecular structure of ChromaLINK Biotin Maleimide

Labeling of proteins with ChromaLINK Biotin eliminates the need to carry out cumbersome and time-consuming HABA assays often employed to quantify biotin incorporation. Instead, biotin incorporation is quantified by means of a simple spectrophotometric measurement at two wavelengths (A280 / A354). Typical labeling results are illustrated in Figure 2 by spectral overlay scans of four samples. As illustrated, bovine Serum Albumin (100 ul @ 1 mg/ml) was labeled at 0, 5, 10, and 20 mole equivalents using ChromaLINK Biotin Maleimide. Spectral analysis illustrates how easy it is to visualize, confirm, and quantify biotin incorporation.

Figure 2. Superimposed spectra of BSA biotinylated using ChromaLINK Biotin Maleimide. Various biotin-to-protein mole equivalents (5X, 10X and 20X) were used. Note the UV-signature at 354nm indicating incorporation of biotin. All spectra were scanned on a Molecular Dynamics SpectraMax PlusTM UV-VIS plate reader (220-420 nm).

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Sulfo ChromaLINK® Biotin (Water Soluble)

Vector Laboratories R&D — Wed, 03 May 2023 04:13:19 +0000

Description

ChromaLINK Biotin contains a UV-traceable chromophore based on ChromaLINK technology to enable reproducibility in your biotinylation process. Now you can measure the degree of biotinylation in minutes, not hours, without the standard curves required for HABA/avidin and fluoro-reporter assays. With a simple, non-destructive UV scan, you can quantify biotin incorporation and ensure reproducible production of consistent batches. This biotinylation reagent is water soluble, which means no DMF or other organic solvent is required to dissolve the compound prior to labeling.

Specifications

Label/Modifier Type	biotin
Reactivity	Streptavidin
Recommended Storage	Desiccated: -15° to -25°C
Applications	Antibody Labeling, Aptamers
Other Name(s)	SoluLINK Bioconjugation

Documents

Citations

Technical Information

Introduction to ChromaLINK Labeling Technology

ChromaLINK Biotin incorporates UV-traceable biotin onto proteins containing lysine residues (amine groups) via a water-soluble-succinimidyl activated ester. ChromaLINK Biotin has been engineered to include many novel features. As illustrated in Figure 1, the molecule’s structure contains a bis-aryl hydrazone chromophore (a), linked by a PEG3 linker arm (b), to biotin (c). This reagent permits direct spectroscopic quantification of incorporated biotin. The extended PEG3 linker preserves biotin/streptavidin affinity and maintains protein solubility after modification while the succinimidyl ester functional group (d), efficiently modifies lysines in aqueous buffers.

Figure 1. Molecular structure of ChromaLINK Biotin (water soluble)

Labeling of proteins with ChromaLINK Biotin eliminates the need to carry out cumbersome and time-consuming HABA assays often employed to quantify biotin incorporation. Instead, biotin incorporation is quantified by means of a simple spectrophotometric measurement at two wavelengths (A280 / A354). Typical labeling results are illustrated in Figure 2 by spectral overlay scans of four samples. As illustrated, bovine IgG (100 ul @ 5 mg/ml) was labeled at 0, 5,10, and 15 mole equivalents using ChromaLINK Biotin. Spectral analysis illustrates how easy it is to visualize, confirm, and quantify biotin incorporation.

Figure 2. Superimposed spectra of bovine IgG biotinylated using ChromaLINK Biotin. Various biotin-to-protein mole equivalents (5X, 10X and 15X) were used. Note the traceable UV-signature @ 354nm indicating incorporation of biotin. All spectra were scanned on a Molecular Dynamics SpectraMax PlusTM UV-VIS plate reader (220-420 nm).

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ChromaLINK® Biotin (DMF Soluble)

Vector Laboratories R&D — Tue, 02 May 2023 21:17:29 +0000

Description

ChromaLINK Biotin contains a UV-traceable chromophore based on ChromaLINK technology, which enables reproducibility in your biotinylation process. Now you can measure the degree of biotinylation in minutes, not hours, without the standard curves required for HABA/avidin and fluoro-reporter assays. With a simple UV scan, you can quantify biotin incorporation and ensure reproducible production of consistent batches.

Specifications

Label/Modifier Type	biotin
Reactivity	Streptavidin
Recommended Storage	Desiccated: -15° to -25°C
Applications	Antibody Labeling, Aptamers
Other Name(s)	SoluLINK Bioconjugation

Documents

Citations

Technical Information

Introduction to ChromaLINK Labeling Technology

ChromaLINK Biotin has been engineered to include many novel features. As illustrated in Figure 1, the molecule’s structure contains a bis-arylhydrazone chromophore (a), linked by a PEG3 linker arm (b), to biotin (c). This reagent permits direct spectroscopic quantification of incorporated biotin. The extended PEG3 linker preserves biotin/streptavidin affinity and maintains protein solubility after modification while the succinimidyl ester functional group (d), efficiently modifies lysines in aqueous buffers.

Figure 1. Molecular structure of ChromaLINK Biotin

Labeling of proteins with ChromaLINK Biotin eliminates the need to carry out cumbersome and time-consuming HABA assays often employed to quantify biotin incorporation. Instead, biotin incorporation is quantified by means of a simple spectrophotometric measurement at two wavelengths (A280 / A354). Typical labeling results are illustrated in Figure 2 by spectral overlay scans of four samples. As illustrated, bovine IgG (100 ul @ 5 mg/ml) was labeled at 0, 5, 10, and 15 mole equivalents using ChromaLINK Biotin. Spectral analysis illustrates how easy it is to visualize, confirm, and quantify biotin incorporation.

Figure 2. Superimposed spectra of bovine IgG biotinylated using ChromaLINK Biotin. Various biotin-to-protein mole equivalents (5X, 10X and 15X) were used. Note the UV-signature at 354nm indicating incorporation of biotin. All spectra were scanned on a Molecular Dynamics SpectraMax PlusTM UV-VIS plate reader (220-420 nm).

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Goat Anti-Biotin, Peroxidase Conjugated

Vector Laboratories R&D — Tue, 02 May 2023 01:10:55 +0000

Description
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Description

Peroxidase conjugated anti-biotin antibodies are produced with the highest specific acitivity enzymes in optimal enzyme/antibody ratios. The covalent linkage between the enzyme and antibody is chosen to provide stable, highly active conjugates for immunohistochemical, in situ hybridization, enzyme immunoassay, or transfer blot applications.

Specifications

Unit Size	1 mg
Applications	Immunohistochemistry / Immunocytochemistry, Immunofluorescence, In situ hybridization, Blotting Applications, Elispot, ELISAs
Concentration	1 mg active conjugate/ml
Recommended Storage	2-8 °C, DO NOT FREEZE
Solution	10 mM sodium phosphate, pH 7.5, 0.15 M NaCl, 0.05% thimerosal
Recommended Usage	Moles peroxidase/mole IgG:3.3 Recommended diluent for microtitration plate assays: 10 mM sodium phosphate, 0.15 M NaCl, pH 7.5, 0.1% Tween 20.Working concentration of HRP anti-biotin is 0.5 – 5.0 µg/ml. Do not use sodium azide with this product since it is an inhibitor of horseradish peroxidase.
Detection Target	Biotin
Conjugate	Peroxidase
Host Species	Goat
Format	Concentrate
Safety Title	WARNING: Cancer – www.P65Warnings.ca.gov

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Citations

Technical Information

Our high affinity polyclonal anti-biotin antibody is produced by hyperimmunizing goats with biotin and is purified by affinity chromatography. Anti-Biotin is available unconjugated, bound to heat stable, cross-linked agarose beads or labeled with enzymes or fluorescein. Agarose bound Anti-Biotin contains 1 mg of Anti-Biotin per ml of settled 4% agarose gel. This product can be used to bind and dissociate biotinylated molecules. Fluorescein labeled Anti-Biotin has been optimally conjugated to provide a bright fluorescent signal. The horseradish peroxidase and alkaline phosphatase conjugates have been produced with the highest specific activity enzymes in optimal enzyme/antibody ratios. The covalent linkage between the enzyme and antibody was chosen to provide stable, highly active conjugates for immunohistochemical, enzyme immunoassay, or transfer blot applications.

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Biotin (Long Arm) Maleimide

Vector Laboratories R&D — Sat, 29 Apr 2023 17:46:04 +0000

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Technical Information

Description

Biotin (Long Arm) Maleimide is designed for biotinylating proteins, nucleic acids or other molecules containing one or more thiol groups. The presence of the 6-aminohexanoate spacer arm between the maleimide group and biotin reduces the possibility of steric hinderance. Molecules to be labeled with Biotin (Long Arm) Maleimide require free thiol groups. In the case of proteins, Biotin (Long Arm) Maleimide will react with exposed cysteine residues. Alternatively, free thiols may be generated by reducing disulfide bonds or by modifying other reactive groups such as primary amines with compounds like Traut’s reagent (2-iminothiolane).

Specifications


Unit Size	12 mg
Target for Labeling	DNA, Oligonucleotides, PNA, Protein, RNA
Recommended Storage	Storage in solution not recommended. – 20°C to – 80 °C.
Molecular Weight	479.6
Required Reactive Group	Thiol Group
Tag/Group Incorporated	Biotin
Safety Title	WARNING: NA – www.P65Warnings.ca.gov

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Citations

Technical Information

Biotin (Long Arm) Maleimide reacts readily under mild acid or neutral pH conditions in aqueous solution with free sulfhydryl groups such as cysteine residues or thiol groups introduced chemically into proteins, nucleic acids, or other macromolecules.

For labeling nucleic acids, thiols can be introduced into DNA, RNA, or oligonucleotides using the 5′ EndTag™ or 3′ EndTag™ nucleic acid labeling systems.

Because of the extraordinary affinity of avidin and streptavidin to biotin and the many biotin-avidin/streptavidin systems available, the biotin label is ideal for a variety of applications including in situ hybridization, blotting, and affinity binding.

The spacer arm in this derivative insures that the biotin is fully accessible to avidin or streptavidin detection systems.

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