General Vitamin B6 ELISA Kit

This competitive Enzyme-Linked Immunosorbent Assay (cELISA) from Antibodies.com is designed for the in vitro quantitative determination of Vitamin B6 in serum, plasma, tissue homogenates and other biological fluids, with reactivity described as Universal. The assay employs a pre-coated 96-well microplate arranged as 12 x 8 well strips, with the plate pre-coated with Vitamin B6 antigen. During incubation, Vitamin B6 present in samples or standards competes with the fixed amount of immobilized antigen for binding sites on a Biotinylated Anti-Vitamin B6 Antibody; consequently, the more analyte present, the less antibody binds to the plate.

Detection is colourimetric, performed via an HRP-Avidin conjugate and a TMB substrate that is catalysed to a blue product and converted to yellow by an acidic stop solution. Absorbance is read at 450 nm in a microplate reader and the analyte concentration is calculated by reference to the standard curve, with the density of yellow being inversely proportional to the amount of Vitamin B6. Performance figures quoted in the manufacturer's specification include an assay range of 3.12-200 ng/ml and a sensitivity of 1.17 ng/ml, with a total assay time of 2h 30m. Spiked recovery testing across five replicates reported average recoveries of 91% (serum, range 80%-102%), 90% (EDTA plasma, range 81%-100%) and 84% (heparin plasma, range 80%-89%), with parallel dilution-linearity results provided at 1:2, 1:4, 1:8 and 1:16 dilutions for each matrix.

The kit is supplied with a pre-coated microplate, lyophilized standard, Detection Solution A, Detection Solution B, a 30X Wash Buffer, Sample Dilution Buffer, TMB Substrate, Stop Solution and adhesive plate sealers. Kit components are stored at +4°C or -20°C as instructed and the kit should not be used past its expiration date.