The HOOK™ Biotin Quant kit estimates biotin incorporation. Biotin labeling is measured using HABA (4’-hydroxyazobenzene-2-carboxylic acid) dye. HABA binds with avidin at biotin binding sites. The HABA-avidin complex produces a characteristic color that absorbs at 500 nm.
In the presence of biotin or biotinylated agents, these agents compete with the HABA for the binding sites. Since biotins have greater affinity for the binding sites, it results in displacement of HABA from the avidin binding sites and proportionate reduction of absorbance. Hence, the amount of biotin present in the solution can be determined by measuring the reduction in absorbance at 500 nm.
Genotech
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