Mouse Surfactant Associated Protein D ELISA Kit

Manufactured by Antibodies.com for the in vitro quantitative determination of mouse Surfactant Protein D/SP-D, this kit from the Assays and Kits range is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) intended for use with serum, plasma, lung lavage fluid, tissue homogenates or other biological fluids. The kit employs the sandwich enzyme immunoassay technique, using an antibody specific for Surfactant Protein D/SP-D that is pre-coated onto a 96-well microtiter plate. Standards and test samples are added to the wells and the target present in each sample is bound by the immobilised antibody. Following incubation, the wells are washed and incubated with a Biotinylated Anti-Surfactant Protein D/SP-D Antibody, which binds the captured protein. HRP-Avidin conjugate is then added, and after further incubation and washing, TMB substrate solution is used to visualise the enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of the acidic stop solution. The density of yellow is proportional to the amount of Surfactant Protein D/SP-D captured in each well. The concentration of the target can be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.

The assay has a detection range of 0.625–40 ng/ml and a sensitivity of 0.285 ng/ml, read on a colorimetric microplate reader, with a total assay time of 4h 30m. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date. Recovery performance has been characterised in serum (n=5, range 80%–102%, average 91%), EDTA plasma (n=5, range 81%–100%, average 90%) and heparin plasma (n=5, range 80%–89%, average 84%). Dilution linearity is reported at 1:2, 1:4, 1:8 and 1:16 dilutions 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%), all at n=5. The platform is a pre-coated microplate arranged as 12 x 8 well strips, with colorimetric detection.