Human Semaphorin 5A ELISA Kit

Antibodies.com offers a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) for the in vitro quantitative determination of human Semaphorin 5A (SEMA5A) in tissue homogenates or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, with an antibody specific for SEMA5A pre-coated onto a 96-well microtiter plate configured as 12 x 8 well strips. Standards and samples are added to the wells, followed by incubation with a Biotinylated Anti-SEMA5A Antibody and an HRP-Avidin conjugate. TMB substrate is then used to visualise the enzymatic reaction, producing a blue colour that changes to yellow upon addition of the acidic stop solution; the density of yellow is proportional to SEMA5A concentration and is read as O.D. absorbance at 450 nm in a microplate reader.

The reported sensitivity is 0.27 ng/ml, with an assay range of 0.781–50 ng/ml. The total assay time is 4 hours 30 minutes. Kit components include a pre-coated 96-well microplate, lyophilized standard (2 vials), Detection Solution A (120 µl), Detection Solution B (120 µl), 30X Wash Buffer (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml) and plate sealers (5 adhesive strips), stored at +4°C or -20°C as instructed.

Reported recovery values (n=5) are: serum average 91% (range 83–100%), EDTA plasma average 95% (range 90–100%) and heparin plasma average 96% (range 91–101%). The product is intended for research use only.