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.
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).
Microplate Path Length Determinator
The path length of the 200-μL sample in the microplate equals the microplate absorbance divided by the cuvette absorbance:
b = VALUE cm.
For Research Use Only. Not for use in diagnostic procedures.