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Biotin Labeling Kits

Download the Protein Labeling & Conjugation Handbook

 

The HOOK™ Biotin kits are designed for the labeling, purification and quantification of proteins with biotin.

 

Protein Labeling

Each kit is supplied with a specific HOOK™ Biotin Reagent that conjugates to proteins through amines, sulfhydryls, carboxyls or carbohydrates.  The amine and sulfhydryl coupling HOOK™ Biotin Reagents couple directly to the protein through their reactive groups, however the carboxyl coupling HOOK™ Biotin Reagents require a carbodiimide crosslinker and the carbohydrate coupling HOOK™ Biotin Reagents require carbohydrate oxidation before coupling.  The HOOK™ Biotin kits include EDC as the carbodiimide crosslinker in the carboxyl coupling kits and sodium meta-periodate for carbohydrate oxidation in the carbohydrate coupling kits.

In addition to the above, each HOOK™ Biotin kit contains a specific Optimizer Buffer that provides the optimal reaction conditions for each HOOK™ Biotin Reagent.

 

Purification

Following the labeling of the protein with the HOOK™ Biotin Reagent the unreacted biotin and other chemicals are rapidly removed from the labeled protein with the supplied SpinOUT™ Columns. These columns use gel filtration to remove the by-products in <10 minutes.

 

Estimation of Biotin Incorporation

Following the purification of the labeled protein, the HOOK™ Biotin kits contain the reagents to estimate biotin incorporation using the HABA and avidin assay. The assay can be performed in <5 minutes.

Simple Antibody Biotinylation   The HOOK™ IgG Biotinylation (Sulfhydryl) kit is designed for the efficient biotinylation of IgG molecules by immobilizing the IgG molecules on a solid support (Figure 1).    IgG molecules contain a histidine rich region in their Fc region and ..
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Simple Antibody Biotinylation   The HOOK™ IgG Biotinylation (Amine) kit is designed for the efficient biotinylation of IgG molecules by immobilizing the IgG molecules on a solid support (see figure 1).   IgG molecules contain a histidine rich region in their Fc region and this fe..
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HOOK™-Biotin-BMMCC reacts with protein free sulhydryl groups. Sulfhydryl reactive reagents are more specific and react only with free sulfhydryl residues (-SH or thiol groups). The side chain of the amino acid cysteine is the most common source of free sulfhydryl groups. If free sulfhydryl res..
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HOOK™-Biotin-Hydrazide reacts with oxidized carbohydrate side chains. Some biotin reagents do not bind directly to the protein itself but conjugate to the carbohydrate residues of glycoproteins. Carbodhydrate reactive biotin reagents contain hydrazides (-NH-NH2) as a reactive group. The hydraz..
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HOOK™-Biotin-LC-Hydrazide reacts with oxidized carbohydrate side chains. Some biotin reagents do not bind directly to the protein itself but conjugate to the carbohydrate residues of glycoproteins. Carbodhydrate reactive biotin reagents contain hydrazides (-NH-NH2) as a reactive group. The hyd..
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HOOK™-Biotin-PDA reacts with protein free sulhydryl groups. Sulfhydryl reactive reagents are more specific and react only with free sulfhydryl residues (-SH or thiol groups). The side chain of the amino acid cysteine is the most common source of free sulfhydryl groups. If free sulfhydryl resid..
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HOOK™-Biotin-PEG2-Amine reacts with protein carboxyl groups in the presence of the heterobifunctional cross-linker EDC. HOOK™-Biotin-Pentylamine, HOOK™-Biotin-PEG2-Amine and HOOK™-Biotin-PEG3-Amine are water soluble, due to its polyethylene glycol (PEG) spacer arm. These agen..
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HOOK™-Biotin-PEG3-Amine reacts with protein carboxyl groups in the presence of the heterobifunctional cross-linker EDC. HOOK™-Biotin-Pentylamine, HOOK™-Biotin-PEG2-Amine and HOOK™-Biotin-PEG3-Amine are water soluble, due to its polyethylene glycol (PEG) spacer arm. These agen..
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HOOK™-Iodoacetyl-LC-Biotin reacts with protein free sulhydryl groups. Sulfhydryl reactive reagents are more specific and react only with free sulfhydryl residues (-SH or thiol groups). The side chain of the amino acid cysteine is the most common source of free sulfhydryl groups. If free sulfhy..
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HOOK™ NHS-Biotin reacts with protein primary amines. Amines, lysine ε-amines and N-terminal α-amines, are the most abundant group in protein molecules and represent the most common target for biotinylation. For example, BSA contains 59 primary amines, of which up to 35 are availa..
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HOOK™-NHS-dPEG™4-Biotin reacts with protein primary amines.  Amines, lysine ε-amines and N-terminal α-amines, are the most abundant group in protein molecules and represent the most common target for biotinylation. For example, BSA contains 59 primary amines, of which ..
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HOOK™-NHS-dPEG™4-Biotin reacts with protein primary amines.  Amines, lysine ε-amines and N-terminal α-amines, are the most abundant group in protein molecules and represent the most common target for biotinylation. For example, BSA contains 59 primary amines, of which ..
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