Streptavidin – VectorLabs

ChromaLINK® Biotin Maleimide

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

Description
Specifications
Documents
Technical Information
Legal Information

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).

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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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).

Legal Information

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Streptavidin, Agarose

Vector Laboratories R&D — Fri, 28 Apr 2023 21:13:05 +0000

Description
Specifications
Documents
Citations
Technical Information

Description

Agarose Streptavidin (SA-5010) can be used to separate biotinylated macromolecules from unbiotinylated materials or for solid phase binding assays.

Features:

Biotin binding activity of the streptavidin optimally preserved through proprietary coupling procedure
Low non-specific binding activity of gel
Protein concentration is 1 mg streptavidin per ml of settled agarose beads
Supplied as a 1:1 suspension in buffer
Binding capacity is approximately 1 mole biotinylated HRP per mole streptavidin

Specifications

Unit Size	2 ml
Applications	Affinity Chromatography
Concentration	1 mg/ml of settled gel
Recommended Storage	2-8 °C, DO NOT FREEZE
Solution	10 mM HEPES, pH 7.5, 0.15 M NaCl, 0.08% sodium azide.
Recommended Usage	Wash gel thoroughly with buffer before use.
Matrix Conjugate	Streptavidin

Documents

Citations

Technical Information

Agarose Streptavidin (SA-5010) is prepared by conjugating streptavidin to heat stable, cross-linked 4% agarose gel beads. To ensure minimal steric interference and low nonspecific binding, streptavidin is conjugated through a hydrophilic spacer arm. The procedure we have developed for coupling streptavidin to agarose preserves the biotin binding activity of the streptavidin. Unlike cyanogen bromide coupling, our procedure does not produce conjugates which can be leached from the gel with solutes such as Tris buffer. Our procedure also does not generate charged groups on the gel that can bind proteins nonspecifically.

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