Biotin ELISA

$680.00

The Eagle Biosciences Biotin ELISA Assay kit is intended for the quantitative determination of Biotin (Vitamin H, Coenzyme R, Vitamin B7) in food products by enzyme linked immunoassay (ELISA).  The Eagle Biosciences Biotin ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

SKU: BIO06-K01 Categories: , ,

Biotin ELISA Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.5 ng/mL
Dynamic Range: 1.0 – 25 ng/mL
Incubation Time: 1.5 hours
Sample Type: Food Products, Cell Culture, Tissue
Sample Size: 3 – 10 g
Controls Included

Product Developed and Manufactured in Germany
Product Support in the USA

Additional Information

Assay 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.

Assay Principle

The Eagle Biosciences Biotin (Vitamin H) 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 labelled 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 Eagle Biosciences Biotin ELISA Assay test sample.

Assay Procedure

  1. Pipet 50 µL standards or prepared samples in duplicate into the appropriate wells of the micro­titer plate. Immediately add 100 µL biotin-AP conjugate into each well.
  2. Cover the microtiter plate with a plastic foil and incubate for 60 minutes at room temperature on a microtiter plate shaker (or 90 minutes without shaker).
  3. Wash the plate three times as follows: Dis­card the contents of the wells (dump or aspi­rate). Pipet 300 µL of diluted washing solu­tion into each well. After the third repetition empty the wells again and remove residual liquid by striking the plate against a paper towel. The wash procedure is critical. In­suf­fi­cient washing will result in poor precision and falsely ele­vated absorbencies.
  4. Pipet 100 µL of substrate solution into each well.
  5. Allow the reaction to develop in the dark (e.g. cup­board or drawer; the chromogen is light-sensitive) for 30 minutes at room tem­perature.
  6. Stop enzyme reaction by adding 100 µL of stop solution (1 M NaOH) into each well. The yellow colour will darken upon addition.
  7. After thorough mixing, measure absorbance at 405 nm (reference wavelength 620 nm), using an ELISA reader. The colour is stable for 30 minutes.

Manual

Product Manual


Publications

Citations

Panneer Selvam, S et. al. Binding of the sphingolipid S1P to hTERT stabilizes telomerase at the nuclear periphery by allosterically mimicking protein phosphorylation. Sci Signal. 2015:8(381) pp.ra58

Related News

Recent Publication in Sci Signal Citing EagleBio’s Biotin ELISA