Rat Aspartate Aminotransferase ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is intended for the in vitro quantitative determination of rat Aspartate Aminotransferase in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.

The assay uses the sandwich enzyme immunoassay technique. An antibody specific for Aspartate Aminotransferase is pre-coated onto a microtiter plate. Standards and test samples are added to the wells, where the analyte is bound by the immobilised antibody. After incubation and washing, a Biotinylated Anti-Aspartate Aminotransferase Antibody is added and binds the captured analyte. An HRP-Avidin conjugate is then added; following further incubation and washing, TMB substrate solution is used to visualise the enzymatic reaction, producing a blue product that changes to yellow upon addition of an acidic stop solution. The density of yellow, read at 450 nm on a colorimetric microplate reader, is proportional to the amount of analyte captured in each well, and concentration is calculated against a standard curve.

Reported sensitivity is 0.117 ng/ml with a product range of 0.312–20 ng/ml. Total assay time is 4h 30m on a pre-coated microplate platform. Sample recovery data are provided: serum range 80%–102% (average 91%, n=5), EDTA plasma range 81%–100% (average 90%, n=5) and heparin plasma range 80%–89% (average 84%, n=5). Linearity of dilution across 1:2, 1:4, 1:8 and 1:16 is reported for serum (87–91%, 87–107%, 74–101%, 92–97%), EDTA plasma (90–105%, 84–101%, 90–101%, 79–108%) and heparin plasma (84–95%, 92–105%, 82–105%, 89–91%), each at n=5. Kit components include a pre-coated microplate, lyophilised standard, Detection Solutions A and B, 30X Wash Buffer, Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers. Components are stored at +4°C or −20°C as instructed, and the kit must not be used past its expiration date.