Porcine Interleukin 1 Alpha ELISA Kit

From Antibodies.com, this Porcine Interleukin 1 Alpha ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of porcine IL-1 alpha in serum, plasma, tissue homogenates, cell lysates, cell culture supernates, or other biological fluids.

The assay employs the sandwich enzyme immunoassay technique. An antibody specific for IL-1 alpha is pre-coated onto a 96-well microtiter plate. Standards and test samples are added to the wells, where IL-1 alpha present in each sample is bound by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-IL-1 alpha Antibody is added, which binds the captured IL-1 alpha. An HRP-Avidin conjugate is then added. After further incubation and washing, TMB substrate solution is used to visualize the HRP enzymatic reaction by catalysis, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of IL-1 alpha captured in each well. Concentration is calculated by reading O.D. absorbance at 450 nm in a microplate reader against the standard curve.

The detection range is 15.6-1000 pg/ml with a sensitivity of <5.6 pg/ml. The assay is read on a colorimetric microplate reader and takes approximately 4h 30m. Reported average recoveries are 91% for serum (n=5, range 80%-102%), 90% for EDTA plasma (n=5, range 81%-100%), and 84% for heparin plasma (n=5, range 80%-89%). Dilution linearity was demonstrated at 1:2, 1:4, 1:8 and 1:16 dilutions across serum, EDTA plasma and heparin plasma. Kit components include a pre-coated 96-well microplate, lyophilized standard, Detection Solutions A and B, 30X wash buffer, sample dilution buffer, TMB substrate, stop solution, and plate sealers. Components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.