Human Caspase 8 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of human Caspase-8 in tissue homogenates, cell lysates, cell culture supernates, or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, with an antibody specific for Caspase-8 pre-coated onto a 96-well microplate configured as 12 x 8 well strips. Standards and test samples are added to the wells, followed by a Biotinylated Anti-Caspase-8 Antibody, HRP-Avidin conjugate, and TMB substrate; the enzymatic reaction is stopped with an acidic stop solution and absorbance is read at 450 nm on a colorimetric microplate reader, with concentration calculated from the standard curve.

Performance characteristics include a product range of 78.1-5000 pg/ml, a sensitivity of <35 pg/ml, and a total assay time of 4 hours 30 minutes. Recovery in serum (n=5) was 80%-102% with an average of 91%; in EDTA plasma (n=5) 81%-100% with an average of 90%; and in heparin plasma (n=5) 80%-89% with an average of 84%. Dilution linearity across 1:2 to 1:16 spanned 87-107% in serum, 79-108% in EDTA plasma, and 82-105% in heparin plasma. Detection is colourmetric on the pre-coated microplate platform.

The kit is suitable for cell lysates, tissue homogenates, and cell culture supernates. Components comprise a pre-coated 96-well microplate, lyophilized standard (2 vials), Detection Solution A (120 µl) and Detection Solution B (120 µl), 30X Wash Buffer (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml), and 5 adhesive plate sealers, with storage at +4°C or -20°C as instructed.