HABA (4'-hydroxyazobenzene-2-carboxylic acid)

Thermo Scientific Pierce HABA is 4'-hydroxyazobenzene-2-carboxylic acid, a simple reagent that enables spectrophotometric (colorimetric) estimation of biotinylation levels of labeled proteins and other molecules. Determine the molar ratio of biotin incorporated into a protein using the HABA-Avidin method. The HABA dye (4'-hydroxyazobenzene-2-carboxylic acid ) binds to avidin to produce a yellow-orange colored complex which absorbs at 500nm. Free biotin will displace the HABA dye and cause the absorbance to decrease. A standard curve can be established using the free biotin to estimate the number of moles of biotin incorporated after biotinylating a protein.

Highlights:

• HABA-avidin complex can be used over a wide range of pH and salt concentrations
• Amount of Avidin can be calculated directly from the increased absorbance at 500nm complexing with the HABA Dye
• Calculate results directly from absorbance values based on extinction coefficients using the procedure outlined in the instructions or use Pierce's online HABA calculator
• Complete kits also available! See Pierce Biotin Quantitation Kit (Part No. 28005) and Fluorescence Biotin Quantitation Kit (Part No. 46610)