Porcine Casp8 (Caspase 8) ELISA Kit

Designed for the sensitive detection of caspase 8 levels in porcine samples, this Sandwich ELISA kit from Assay Genie enables precise quantification in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Caspase 8 is a key enzyme in the initiation of apoptosis, playing a crucial role in cell death pathways, with dysregulation implicated in cancer, autoimmune disorders, and infectious diseases. The kit uses a colorimetric double-antibody sandwich detection method, with a reported sensitivity of 46.875 pg/mL and a working range of 78.125–5000 pg/mL.

The assay procedure follows six steps: equilibrate reagents and TMB substrate to room temperature and set up standards, samples and controls; incubate at 37°C for 90 minutes; add biotin-labelled detection antibody and incubate at 37°C for 60 minutes; add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes; add TMB substrate and incubate in the dark for 10–20 minutes; and add stop solution before reading absorbance at 450 nm. Sample preparation covers serum (allow blood to clot, centrifuge at 1000 × g for 20 minutes), plasma (collect using anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C), tissue homogenate (homogenise in PBS with protease inhibitors), cell culture supernatant (centrifuge at 2500 rpm for 5 minutes), and cell lysate (use lysis buffer with protease inhibitors).

Components supplied include a dismountable pre-coated ELISA microplate, lyophilised standard, concentrated biotin-labelled antibody, HRP-Streptavidin conjugate, TMB substrate, sample dilution buffer, antibody dilution buffer, SABC dilution buffer, stop solution, wash buffer, plate sealer, and technical manual. Storage is at 2–8°C for 12 months, with the standard and plate strips additionally storable at -20°C. Linearity and recovery have been characterised in serum, EDTA plasma and heparin plasma matrices.