Human Semaphorin 3A ELISA Kit

This Antibodies.com human Semaphorin 3A ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) for the in vitro quantitative determination of human Semaphorin 3A in serum, plasma, tissue homogenates, and other biological fluids. The assay uses a pre-coated 96-well microtiter plate with colorimetric detection at 450 nm, a total run time of 4h 30m, and is read on a colorimetric microplate reader. The kit has a reported sensitivity of <0.57 ng/ml and a measuring range of 1.56-100 ng/ml.

In the assay workflow, standards and samples are added to wells where Semaphorin 3A binds the immobilized capture antibody. After washing, a Biotinylated Anti-Semaphorin 3A Antibody is added, followed by HRP-Avidin conjugate. TMB substrate then visualises the HRP enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution. The intensity of the yellow colour is directly proportional to the amount of Semaphorin 3A captured, allowing concentration to be calculated from the standard curve.

Sample recovery was validated across serum, EDTA plasma, and heparin plasma (each n=5), with average recoveries of 91%, 95%, and 94% respectively, and dilution linearity confirmed across 1:2, 1:4, 1:8, and 1:16 sample dilutions. Kit components include a pre-coated 96-well microplate, lyophilized standard, biotinylated detection reagents, HRP conjugate, wash buffer, sample dilution buffer, TMB substrate, stop solution, and plate sealers, stored at +4°C or -20°C as instructed.