Rat Caspase 3 ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of rat Caspase-3 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. The assay employs a 96-well microtiter plate pre-coated with an antibody specific for Caspase-3, and the absorbance is read at 450 nm on a colorimetric microplate reader.

Standards and test samples are added to the wells, where Caspase-3 is captured by the immobilised antibody. After washing, a biotinylated anti-Caspase-3 antibody is added, followed by HRP-Avidin conjugate and TMB substrate. The HRP-catalysed reaction produces a blue product that turns yellow upon addition of acidic stop solution, with the intensity of yellow proportional to the amount of Caspase-3 captured in each well. The stated product range is 1.56–100 ng/ml, with a sensitivity of <0.49 ng/ml, and total assay time is 4 hours 30 minutes. Kit components should be stored at +4°C or –20°C as instructed, and the kit should not be used past its expiration date.

Reported recovery values are 80–102% (average 91%) in serum (n=5), 81–100% (average 90%) in EDTA plasma, and 80–89% (average 84%) in heparin plasma, with sample dilution linearity characterised at 1:2, 1:4, 1:8 and 1:16 for each matrix. The kit includes a pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard (2 vials), detection solutions A and B (120 µl each), 30X wash buffer (20 ml), sample dilution buffer (45 ml), TMB substrate (9 ml), stop solution (6 ml), and plate sealers (5 adhesive strips).