Human Phospholipase A2, Group IID ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (ELISA) from Antibodies.com is intended for the in vitro quantitative determination of human SPLASH in serum, plasma, tissue homogenates or other biological fluids. The 96-well microplate is pre-coated with an antibody specific for SPLASH. Standards and test samples are added to the wells and the analyte present is captured by the immobilized antibody. After washing, a Biotinylated Anti-SPLASH Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the HRP enzymatic reaction, producing a blue product that turns yellow on addition of the acidic stop solution. Absorbance is read at 450 nm on a colorimetric microplate reader and the SPLASH concentration is calculated against the standard curve.

Assay sensitivity is <0.32 ng/ml, with a measuring range of 0.937–60 ng/ml. In spike-and-recovery testing, average recoveries were 91% in serum (n=5, range 83–100%), 94% in EDTA plasma (n=5, range 90–98%) and 94% in heparin plasma (n=5, range 90–99%). Linearity has been demonstrated at sample dilutions of 1:2, 1:4, 1:8 and 1:16 across serum, EDTA plasma and heparin plasma samples (n=5 per matrix). Total assay time is 4 hours 30 minutes.

Kit components include a pre-coated 96-well microplate, lyophilised standard, Detection Solution A, Detection Solution 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 should not be used past its expiration date.