Vitamin H (Biotin) ELISA

Enzyme immunoassay for the quantitative determination of Biotin (Vitamin H) in food.

Background: Biotin serves as the prosthetic group of enzymes, which catalyze carboxylations in the organism. For this purpose, biotin is bound via its carboxy group to lysin residues of carboxylases, and the transfer of carbon dioxide takes place after its attachment to a nitrogen atom of biotin, forming the so-called active carbon dioxide.
The awareness of the population for a good health and its interest in healthy nutrition has increased significantly during the last years. After the content of vitamins in his nourishment has gained importance for the consumer, food has partially been vitaminized by the manufacturer. When there exists a lack of biotin, seborrhoea, dermatitis, anorexia, muscle pain, tiredness and nervous disorders can appear. As biotin is synthesized by the human intestinal flora, deficiency symptoms are rare, appear however after excessive ingestion of raw egg white, which can be explained by its content of biotin-binding avidin.

Description: The Vitamin H (Biotin) quantitative test is based on the principle of the enzyme linked immunosorbent assay. Avidin, which shows a high affinity to biotin, is bound on the surface of a microtiter plate. Biotin containing samples or standards and a biotin-alkaline phosphatase conjugate are given into the wells of the microtiter plate. Enzyme labeled and free biotin compete for the binding sites. After one hour incubation at room temperature, the wells are washed with diluted washing solution to remove unbound material. A substrate solution is added and incubated for 30 minutes, resulting in the development of a yellow colour. The colour development is inhibited by the addition of a stop solution. The yellow colour is measured photometrically at 405 nm. The concentration of biotin is indirectly proportional to the colour intensity of the test sample.

Product Features:
- The kit contains reagents for 96 determinations;
- Microtiter plate consisting of 12 strips with 8 breakable wells;
- Biotin Standards: 0, 1, 2.5, 5, 10, 25 ng/mL;
- ELISA reader at 405 nm;
- Standard range: 1- 25 ng/mL;
- Sensitivity analytical: 0.5 ng/mL.