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After three washes with TBST buffer, the membranes were incubated at 4 C for 4 h in TBST containing the purified His-tag protein at a final concentration of 35 g/mL

Once activated, caspase-3 initiates the cleavage of a downstream substrate, such as nuclear poly (ADP-ribose) polymerase (PARP), which facilitates cellular disassembly and serves as a biomarker of cell apoptosis