Necroptosis Inducer Kit

The Necroptosis Inducer Kit provides researchers a convenient approach to induce programmed necrosis (necroptosis), a regulated cell death pathway triggered by tumor necrosis factor (TNF)-receptor or Toll-like receptor signaling under caspase-inhibited conditions. Necroptosis induction involves phosphorylation and activation of kinases RIP1 and RIP3, ultimately leading to necrotic cell death. This ready-to-use reagent mixture combines recombinant human TNF-α, a cytokine initiating necroptotic signaling; SM-164, a Smac mimetic facilitating TNF-α-mediated apoptotic pathways; and Z-VAD-FMK, a cell-permeable pan-caspase inhibitor preventing apoptosis. Researchers commonly employ this kit in tumor biology, neurodegenerative disease, and inflammation studies. For bacterial transformations, chemically competent cells (DH5α strain; genotype: fhuA2 Δ(lacZ-argF)U169 phoA glnV44 Φ80ΔlacZ ΔM15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17) with ≥10^8 cfu/μg transformation efficiency are recommended. Antibiotic selection is achieved using ampicillin solution (100 mg/mL stock, final concentration 100 μg/mL). For mammalian cell cryopreservation, cell freezing medium achieves cell recovery rates ≥90%. Cell transfection reagents included provide ≥80% efficiency across various commonly used cell lines such as HEK293 and HeLa.