Phospho-MLKL-S345 Rabbit mAb for WB, IP, ELISA - Q9D2Y4

Core Product Specifications and Parameters

Biological Background: Mixed lineage kinase domain-like protein Function and Localization

• Mixed lineage kinase domain-like protein (MLKL) is a pseudokinase critical for necroptotic cell death. An alternative splicing isoform may exist.
• MLKL is the terminal effector of TNF-induced necroptosis. Upon phosphorylation by RIPK3, it forms homotrimers, translocates to the plasma membrane, and triggers calcium influx and plasma membrane rupture, executing programmed necrosis. Related references: PMID:23835476, PMID:24012422, PMID:24019532
• Despite its kinase-like domain, MLKL lacks protein kinase activity and functions as a pseudokinase. Related references: PMID:24012422
• In response to orthomyxovirus infection, nuclear MLKL can be activated by ZBP1 and RIPK3, leading to disruption of the nuclear envelope and leakage of immunogenic DNA into the cytosol. Related references: PMID:32200799, PMID:32296175
• MLKL is predominantly cytoplasmic but can associate with the cell membrane and nucleus depending on activation state.
• MLKL is highly expressed in the thymus, colon, intestine, liver, spleen, and lung, with lower levels in skeletal muscle, heart, and kidney, and is absent in brain.
• MLKL contains a coiled-coil domain and an ATP-binding site, although it does not exhibit kinase activity. It is a phosphoprotein regulated by multiple phosphorylation events.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phospho-peptide corresponding to mouse MLKL S441; confirm antibody specificity by treating samples with lambda phosphatase or using non-phosphorylated peptide competition.
• For Western blot, a band around 54 kDa is expected. Use appropriate positive controls such as necroptotic stimulus-treated mouse cell lysates (e.g., TNF+ZVAD+SMAC mimetic).
• In immunoprecipitation, pre-clearing and optimized antibody:lysate ratios are recommended to reduce background.
• ELISA performance may depend on coating conditions; test both direct and sandwich ELISA formats.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers and research labs with comprehensive raw material solutions for necroptosis and cell death research. We offer validated antibody pairs, recombinant proteins, and ancillary reagents to support assay development. For high-demand targets like MLKL, our sourcing network ensures bulk availability and consistent quality, reducing your development timeline.