BIOTIN (VITAMIN H) ELISA KIT
INTENDED USE
The Biotin (Vitamin H) ELISA Kit Assay is intended for the quantitative determination of Biotin (Vitamin H) in serum, plasma, drugs and foods. For research use only.
INTRODUCTION
In the late 1950s Lynen and Wakil reported the discovery of covalently bound biotin with a coenzyme function. Biotin is a growth factor present in minute amounts in every living cell. It plays an indispensable role in numerous naturally occurring carboxylation reactions. The daily biotin requirements for adults lie between 100 and 200 mg. Renewed interest in the role of biotin in human nutrition and therapy stems from the accumulating reports of various biotin-responsive syndromes. A variety of methods for the assay of biotin are currently available. However most of them are either cumbersome to perform or based on the use of radioactive materials. This assay is a convenient reciprocal assay for biotin. It is extremely sensitive, easy to perform and doesn’t use radiolabels.
PRINCIPLE OF THE TEST
Biotin (Vitamin H) ELISA Kit
Biotin (Vitamin H) ELISA Kit Assay is intended for the quantitative determination of Biotin (Vitamin H) in serum, plasma, drugs and foods. The Biotin (Vitamin H) ELISA Kit test is a competitive enzyme-linked assay for biotin determination in human serum. Samples, standards and controls are incubated with streptavidin-enzyme conjugate for 15 minutes at 18-25 oC. Then 100 μl of each incubation mixture is transferred in duplicate into microtiter plate wells coated with biotin-albumin conjugate. After a second incubation step of 30 minutes at 18-25 oC, the enzyme substrate TMB is added. Finally, the reaction is terminated by an acidic stop solution causing a color change from blue to yellow. The color intensity is inversely proportional to the biotin concentration. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standards. Biotin present in the patient samples is determined from this curve.
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