Biotin Quantitation Kit (Colorimetric) (BN01027)

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  • Biotin Quantitation Kit (Colorimetric) (BN01027)
  • Biotin Quantitation Kit (Colorimetric) (BN01027)


ELISA Kit Technical ManualMSDS

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.

Assay Genie offers a variety of ready-to-use Biotinylation reagents. It is often desired to determine the degree of substitution of such biotinylated biomolecules. Assay Genie'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.

Figure: (a) Biotin Standard Curve (b) Linear plot obtained for the amount of biotin (nmol) vs amount of biotinylated BSA. Assays were   performed according to the kit protocol.

Key Information Description

Product SKU


Kit Size

100 assays


Absorbance (500 nm)


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


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

Kit Contents

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




Gel Pack

Research Use

For Research Use Only! Not For Use in Humans.

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