Thermo Scientific Pierce Biotin Quantitation Kit includes aliquots of HABA reagent and biotinylated protein standard for convenient colorimetric determination of biotinylation levels in labeled antibodies and other proteins.
Features of the Biotin Quantitation Kit:
• Colorimetric – requires a plate reader or spectrophotometer set to measure at 500nm • Reliable – proven, well-characterized method for determination of biotinylation levels • Convenient – kit provides the essential reagents and standards to perform the assay with ease • Flexible – adaptable to spectrophotometer cuvettes or standard plate readers with 96-well microplates • Options – calculate results directly from absorbance values based on extinction coefficients using the procedure outlined in the instructions • Robust – supplied HABA-avidin complex can be used over a wide range of pH and salt concentrations
HABA dye (4'-hydroxyazobenzene-2-carboxylic acid) is a reagent that enables quick estimation of the mole-to-mole ratio of biotin to protein in a solution. The Pierce Biotin Quantitation Kit contains a premix of HABA and avidin and a biotinylated horseradish peroxidase (HRP) positive control. The HABA-Avidin Premix is supplied in convenient Thermo Scientific No-Weigh Microtube packaging, which eliminates the difficulties associated with weighing small quantities of reagent.
To quantitate biotinylation, a solution containing the biotinylated protein is added to a mixture of HABA and avidin. Because of its higher affinity for avidin, biotin displaces the HABA and the absorbance at 500nm decreases proportionately. By this method, an unknown amount of biotin present in a solution can be quantitated in a single cuvette by measuring the absorbance of the HABA-avidin solution before and after addition of the biotin-containing sample. The change in absorbance relates to the amount of biotin in the sample by the extinction coefficient of the HABA-avidin complex. View online HABA Calculator.
For Research Use Only. Not for use in diagnostic procedures.