Biotin Quantitation Kit (Colorimetric)

Biotin (Vitamin H) is an important biomolecule that has been widely used in Biotinylation reactions to label antibodies or other proteins of interest. Due to its low molecular weight, biotin usually does not cause any significant changes in protein conformation and biological activity. Biotinylated proteins/antibodies are widely isolated and assayed with streptavidin pull-down assay, affinity chromatography, ELISA and Western blotting etc. Biovision offers a variety of ready-to-use Biotinylation reagents (Cat. # 2346, 2345, 2347, 2322, 2326, 2323). It is often desired to determine the degree of substitution of such biotinylated biomolecules. Biovision’s Biotin Assay Kit is based on the differential binding of Streptavidin to Biotin and a dye, 2-(4-hydroxyazobenzene) benzoic acid (HABA). When HABA is bound to Streptavidin, addition of free biotin or biotinylated biomolecules results in its displacement in the reaction. These changes are manifested in terms of decrease in the overall absorption (OD 500 nm) and can be quantitatively correlated to the amount of biotin present in solution. The kit also provides Biotin as a Standard and Biotinylated BSA as a positive control. This kit can be used to estimate as low as 60 pmol or 15 ng of biotin in solution.

• Detection method: Absorbance (500 nm) • Application: Measuring free biotin or the substitution degree of Biotinylation (biotin labels) of proteins and antibodies

• Simple & convenient • This kit can be used to estimate as low as 60 pmol or 15 ng of biotin in solution

Biotin

Vitamin H Quantitation Kit (Colorimetric)

• Biotin Assay Buffer • Biotin Assay Reagent A • Biotin Assay Reagent B • Biotin Standard Solution (10 mM in DMSO) • Biotinylated BSA (2 mg/ml)

-20°C

gel pack

12 months

see datasheet

Measuring free biotin or the substitution degree of Biotinylation (biotin labels) of proteins and antibodies

Biotin conjugates can be detected by horseradish peroxidase, alkaline phosphatase substrates or anti biotin conjugated antibodies. Avidin and Streptavidin bind to the small biotin and are couple to HRP or AP for ELISA. To break the streptavidin Biotin bond we suggest to use a 6 molar guanidine HCl solution with acidity of pH 1.6.Colorimetric assays or detection use UV absorption or enzymatic color reaction.

Biotinylated