Biotin Assay Kit (Colorimetric) (ab185441)
Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Purified protein
- Sensitivity: 15 ng
Overview
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Product name
Biotin Assay Kit (Colorimetric) -
Detection method
Colorimetric -
Sample type
Purified protein -
Assay type
Semi-quantitative -
Sensitivity
15 ng -
Product overview
Abcam’s Biotin Quantitation Kit (Colorimetric) (ab185441) 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.
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Notes
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. It is often desired to determine the degree of substitution of such biotinylated biomolecules.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Biotin Assay Buffer 1 x 25ml Biotin Assay Reagent A 1 x 8ml Biotin Assay Reagent B 1 x 1ml Biotin Standard Solution (10 mM in DMSO) 1 x 100µl Biotinylated BSA (2 mg/ml) 1 x 200µl -
Research areas
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Relevance
Biotin is a water soluble vitamin, generally classified as a B complex vitamin, also called vitamin B4. After the initial discovery of biotin, nearly forty years of research were required to establish it as a vitamin. Biotin is required by all organisms but can only be synthesized by bacteria, yeasts, molds, algae, and some plant species. Biotin is required as prosthetic group of enzymes involved in incorporation of carbon dioxide into organic compounds. Biotin has a MW of 244 Da. -
Alternative names
- Vitamin B4
Images
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Biotin Standard Curve
This is example data only.
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Amount of biotin vs amount of biotinylated BSALinear plot obtained for the amount of biotin (nmol) vs amount of biotinylated BSA. Assays were performed according to the kit protocol. This is example data only.