Human Adenylate Cyclase 5 ELISA Kit

Manufactured by Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human adenylate cyclase 5 (AC5) in tissue homogenates, cell lysates, or other biological fluids. The kit employs the sandwich enzyme immunoassay technique using a pre-coated 96-well microplate arranged as 12 x 8 well strips, with an antibody specific for AC5 immobilised on the wells. Standards and test samples are added to the wells so that AC5 present in each sample is captured by the immobilised antibody.

Following incubation and washing, a Biotinylated Anti-AC5 Antibody is added, which binds the captured AC5 in each well. An HRP-Avidin conjugate is then added, and TMB substrate solution is used to visualise the enzymatic reaction, producing a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of AC5 captured, and the concentration is calculated by reading the O.D. absorbance at 450nm in a colorimetric microplate reader against the standard curve. The assay has a working range of 0.312-20 ng/ml and a sensitivity of 0.124 ng/ml, with a total assay time of 4h 30m.

Validation data from the manufacturer report recovery in serum (range 80-102%, average 91%), EDTA plasma (range 81-100%, average 90%), and heparin plasma (range 80-89%, average 84%) at n=5, with dilution linearity demonstrated at 1:2, 1:4, 1:8, and 1:16 across the same sample types. Kit components include a pre-coated microplate, lyophilised standard, detection solutions A and B, wash buffer, sample dilution buffer, TMB substrate, stop solution, and adhesive plate sealers, to be stored at +4°C or -20°C as instructed and not used past the stated expiration date.