| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Food, cosmetics, supplements, and biotinylated proteins or antibodies |
| Shipping | |
| Species | |
| Storage |
Overview
For quantitative determination of biotin in food, cosmetic, and biotinylated proteins. The assay uses OD500nm for signal readout. Compatible sample input includes Food, cosmetics, supplements, and biotinylated proteins or antibodies. Typical stated assay timing is 20 min.
Key elements and design rationale
- Readout format: OD500nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Food, cosmetics, supplements, and biotinylated proteins or antibodies, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 8 µM for interpreting low-signal samples.
- Feature emphasis: 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).
Additional feature notes highlight 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. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of biotin within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
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.
Detection method
Colorimetric (OD 500 nm).
Detection limit and analytical sensitivity
Reported detection limit: 8 µM.
Procedures and timing
Stated procedure or timing information: 20 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- Short assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.
Common research applications
- Quantify biotin in food, cosmetics, supplements by OD500 nm readout.
- Compare treatment or phenotype groups using matched food, cosmetics, supplements handling.
- Monitor time-course or pre/post changes in food, cosmetics, supplements across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
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 laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
