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