Human Cleaved CASP9 (Cleaved Caspase-9) ELISA Kit (HUFI04740)

The cleaved (activated) form of caspase-9 is the biochemical marker of intrinsic apoptotic pathway engagement, since processing of the zymogen at Asp315 by the apoptosome generates the catalytically competent p35 fragment that propagates the downstream caspase cascade. This quantitative immunoassay from Assay Genie uses monoclonal capture antibodies directed against that Asp315 neoepitope, so it selectively reports the activated fragment rather than full-length pro-caspase-9 protein. That selectivity matters in experiments where total caspase-9 levels remain flat while activation status changes dramatically, a common situation during chemotherapy exposure, neurotrophic factor withdrawal, ischemic insult, or developmental cell death in human cell culture and tissue models.

The kit ships as a complete research-use package containing pre-coated plate wells, a matched biotinylated detection antibody, an enzyme conjugate, calibrated recombinant standards, wash buffer, substrate solution, and stop reagent, with reagents supplied for 96 Assays run in parallel. The workflow is performed at room temperature using standard immunoassay incubation and wash steps, and the absorbance-based endpoint is quantified against the supplied standard curve to yield interpolated sample concentrations appropriate for comparing treatment groups or time points within a single study.

Typical applications include monitoring apoptosis kinetics in cultured human cell lines, quantifying pathway activation in disease-relevant primary samples, screening small molecules or biologics for pro- or anti-apoptotic activity, and validating CRISPR or RNAi knock-down phenotypes that target the intrinsic death pathway. Because the antibody pair was raised against the human neoepitope, results from non-human species should be interpreted cautiously and confirmed by an orthogonal method such as Western blot probing the cleaved fragment.