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     All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.    

  
  

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     Western blot analysis of Phospho-MLKL (S358) expression in (1) HT-29 cell lysate; (2) HT-29 cell lysate treated with TNF alpha+ Smac mimetic+ z-VAD.    

  
  

The Phospho-MLKL(S358) antibody is a specialized tool used to detect phosphorylation of the mixed lineage kinase domain-like (MLKL) protein at serine 358 (S358), a critical post-translational modification in necroptosis—a form of programmed necrotic cell death. MLKL acts as the terminal executor of necroptosis, transitioning from an inert state to an active oligomerized form upon phosphorylation by receptor-interacting protein kinase 3 (RIPK3). This phosphorylation event, particularly at S358 (human; equivalent to murine S345), triggers MLKL's conformational change, enabling its translocation to the plasma membrane to induce membrane permeabilization and cell death.

The antibody is widely employed in research to study necroptotic signaling in physiological and pathological contexts, including inflammatory diseases, neurodegenerative disorders, and cancer. By specifically recognizing phosphorylated S358. it allows detection of activated MLKL via techniques like Western blotting, immunofluorescence, or immunohistochemistry. Validating its specificity is crucial, often achieved using MLKL-knockout cell lines or RIPK3/MLKL inhibitors to confirm signal dependency. Researchers rely on this tool to dissect regulatory mechanisms, screen therapeutic agents targeting necroptosis, or explore disease models where dysregulated cell death pathways contribute to pathogenesis. Its application bridges mechanistic insights into cell death control and potential clinical interventions.