HABA is a dye with absorbance and avidin-binding properties that help enable the quantitation of biotin in a sample. When HABA is bound to avidin, the complex absorbs light at 500 nm (e = 34,000/M cm). When a biotin-containing sample is added, the biotin binds strongly to avidin and displaces the weakly bound HABA resulting in a decrease in absorbance relative to the amount of HABA displaced.

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This HABA calculator performs the computations necessary to determine the average number of biotin groups per protein molecule (i.e., the biotin:protein molar ratio) following a biotin-labeling reaction. You will need to supply the measured absorbance values of the HABA-avidin solution before and after addition of the biotin sample as well as information about the biotinylated protein (i.e., molecular weight and concentration in milligrams per milliliter). For a detailed explanation of the calculations, consult the instructions for Thermo Scientific Pierce HABA Reagent (Catalog number 28010) or the Thermo Scientific Pierce Biotin Quantitation Kit (Catalog number 28005).

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Choose cuvette format when using a microplate with a reader that automatically corrects for path length or when using a spectrophotometer device having a fixed path length.