Anti-MLKL Rabbit Monoclonal Antibody for WB, IF/ICC, IHC-P, ELISA - Q8NB16

Core Product Specifications and Parameters

Biological Background: MLKL Function and Localization

• MLKL is a pseudokinase that lacks intrinsic protein kinase activity, functioning as a critical executor of TNF-induced necroptotic cell death. Related references: PMID:22265413, PMID:22265414, PMID:22421439

• Upon phosphorylation by RIPK3, MLKL undergoes homotrimerization and translocates to the plasma membrane, where it triggers calcium influx and plasma membrane rupture, hallmark features of necroptosis. Related references: PMID:22265413, PMID:22265414, PMID:22421439

• In addition to cytoplasmic necroptosis, MLKL can also promote nuclear necroptosis during orthomyxovirus infection. ZBP1 senses viral Z-RNA and activates RIPK3 in the nucleus, leading to MLKL phosphorylation, nuclear envelope disruption, and cytosolic leakage of DNA (By similarity).

• MLKL binding to inositolhexakisphosphate (InsP6) is essential for its necroptotic function, linking phosphoinositide signaling to cell death execution. Related references: PMID:29883610

• Subcellularly, MLKL resides primarily in the cytoplasm but shuttles to the cell membrane and nucleus upon activation, consistent with its dual roles in membrane and nuclear envelope disruption.

• The protein contains an N-terminal coiled-coil domain and a C-terminal pseudokinase domain, and is subject to alternative splicing, yielding isoforms that may regulate necroptosis or have unknown functions.

• Post-translational modifications, particularly RIPK3-mediated phosphorylation, are critical for MLKL activation, oligomerization, and membrane targeting; other modifications and interactions may fine-tune its activity.

Experimental Guidance and Technical Tips

• The immunogen used spans amino acids 369-471 within the C-terminal pseudokinase domain of human MLKL (NP_689862.1). This region is likely to recognize both the full-length protein and any isoforms containing this segment; confirm specificity in your model system by RNAi or knockout controls.

• For Western blot, a band around 54kDa is expected for human MLKL. Optimal dilution should be empirically determined using necroptosis-induced cell lysates (e.g., TNF-alpha + Smac mimetic + z-VAD-fmk treatment) as positive controls.

• IF/ICC and IHC-P applications may require antigen retrieval and permeabilization optimized for your sample type. Necroptotic stimuli can enrich MLKL at the plasma membrane, providing a dynamic localization control.

• ELISA protocols can be developed using recombinant MLKL protein or cell lysates; consider paired antibody sets for sandwich ELISA to enhance sensitivity.

• Cross-reactivity is reported against human and mouse; however, validation in the relevant species and tissue is recommended before large-scale studies.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is your integrated partner for IVD raw material sourcing, offering validated antibody pairs, optimized monoclonal/polyclonal antibodies, bulk ancillary reagents, and rare-target procurement to support your diagnostic development from concept to clinic. For necroptosis and cell death research, we provide a reliable supply of anti-MLKL antibodies and complementary reagents to accelerate assay development and biomarker discovery.