QuantiChrom™ Biotin Assay Kit
- Quantitative determination of biotin in food, cosmetic, and biotinylated proteins.
- Fast and sensitive. Linear detection range: 8 to 200 µM biotin with 30 µL sample (96-well) or 10 to 200 µM biotin with 10 µL sample (384-well).
- Convenient. The procedure involves adding a single working reagent and reading after 20 minutes.
- High-throughput. "Add-mix-read" type assay. Can be readily automated as a high-throughput 96-well or 384-well plate assay for thousands of samples per day.
- Food, cosmetics, supplements, and biotinylated proteins or antibodies
- 100 tests (96-well), 320 tests (384-well)
- 8 μM
- 12 months
- BIOTIN , or Vitamin B7, is a water soluble vitamin involved in metabolism, cell growth, and protein synthesis. It is a cofactor to multiple carboxylases necessary for metabolizing fatty acids, glucose, and amino acids. Biotin is found in a wide range of foods and is often taken as a dietary supplement. In the biotechnology industry, biotin is commonly conjugated to proteins in a process called biotinylation. These biotinylated proteins can then be specifically selected using streptavidin and/or avidin's strong affinity for biotin in biochemical assays such as ELISAs.
BioAssay Systems' Biotin Assay Kit is based on avidin's weak affinity for 4'-hydroxyazobenzene-2-carboxylic acid (HABA) and strong affinity for biotin. The avidin-HABA complex reagent has an absorbance at 500nm. When the colored avidin-HABA reagent is introduced to biotin, the biotin binds to avidin, displacing the HABA and causing a decrease in absorbance. The decrease in absorbance at 500nm is directly proportional to the biotin in the sample.
The blank value is higher than the standard values. What is wrong?This is completely normal. The assay is measuring the decrease in absorbance at 500nm. The higher the concentration of biotin, the lower the absorbance will be in the calculation.
My sample is reporting much lower levels of biotin than I was expecting?The biotin levels in your sample may be significantly greater than the kit detection limit. We recommend diluting your sample and assaying again. You may wish to perform a serial dilution on the sample to determine the optimal dilution factor. Aim for a dilution that yields a concentration read out somewhere in the middle of the standard curve. Correct for the dilution factor in the calculation.
Some of my samples are getting negative concentration read outs?This is likely indicative no biotin or biotin below the kit detection limit.
The kit arrived thawed and at room temperature, but the protocol says to store it at -20°C. Is the kit still ok to use?Yes, the kit is shipped at room temperature and is stable for the duration of shipment at that temperature. Upon receipt, the kit should be stored at -20°C.
For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.