Mouse Calreticulin ELISA Kit

Antibodies.com Mouse Calreticulin ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of mouse Calreticulin in serum, plasma, tissue homogenates, or other biological fluids. The assay employs a pre-coated 96-well microtiter plate configured as 12 x 8 well strips, onto which an antibody specific for Calreticulin has been immobilised. Standards and test samples are added to the wells and Calreticulin present in each sample is captured by the immobilised antibody. After washing, a biotinylated anti-Calreticulin antibody is added, followed by an HRP-avidin conjugate and further incubation. TMB substrate solution is then used to visualise the HRP enzymatic reaction by catalysis, producing a blue-coloured product that changes to yellow after addition of acidic stop solution. Absorbance is read at 450 nm in a colorimetric microplate reader and the concentration of Calreticulin is calculated by reference to the standard curve.

The assay has a sensitivity of <0.067 ng/ml and a standard curve range of 0.156–10 ng/ml, with a total assay time of 4 hours 30 minutes. Recovery was assessed in serum (range 80%–102%, average 91%, n=5), EDTA plasma (81%–100%, average 90%, n=5) and heparin plasma (80%–89%, average 84%, n=5). Dilution linearity was evaluated at 1:2, 1:4, 1:8 and 1:16 in serum, EDTA plasma and heparin plasma. Detection is by colorimetric measurement at OD450.

Kit components include a pre-coated 96-well microplate, lyophilised standard, detection solution A, detection solution B, wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers. Kit components should be stored at +4°C or -20°C as instructed in the protocol, and the kit should not be used past its expiration date.