Mixed lineage kinase domain-like protein (UniProt: Q8NB16; also known as hMLKL) is encoded by the MLKL gene (Gene ID: 197259) in human. MLKL is a pseudokinase that is a terminal protein in the pro-inflammatory necroptotic cell death program. It plays a critical role in TNF induced necroptosis. Although it contains a protein kinase domain (aa 194-469), it lacks several residues that are essential for protein kinase activity. It is activated following phosphorylation at Thr357/Ser358 by RIPK3, which leads to its homotrimerization and localization to the plasma membrane where it binds to highly phosphorylated inositol (InsP6) and cardiolipin and disrupts membrane integrity to cause necrotic cell death. Under basal conditions, MLKL is shown to reside in small puncta that are distributed evenly throughout the cytoplasm, but it is detected in plasma membrane shortly after its phosphorylation. It contains two coiled coil regions (aa 55-84 and 139-180). The second coiled coil region is responsible for its oligomerization. MLKL is also reported to be involved in endosomal trafficking, independently of induction of necroptosis, whereby it enhances degradation of receptors and ligands. Two isoforms of MLKL have been described that are produced by alternative splicing. (Ref.: Samson, AL., et al. (2020). Nat. Commun. 11(1); 3151; Yoon, S., et al. (2017). Immunity. 47(1); 51-65; Wang, H., et al (2014). Mol. Cell 54(1); 133-146).
Clone 10C2 is a rat monoclonal antibody that detects human Mixed lineage kinase domain-like protein (MLKL). It targets an epitope within the C-terminal region.
GST-tagged recombinant fragment corresponding to 282 amino acids from human Mixed lineage kinase domain-like protein (MLKL) expressed in baculovirus expression system.
Quality Control Testing
Evaluated by Western Blotting in wild-type HT-29 cell lysate.
Western Blotting Analysis (WB): A 1:500 dilution of this antibody detected MLKL in lysate from wild-type HT-29 cells, but not in MLKL knockout HT-29 cells.
Immunocytochemistry Analysis: A representative lot detected MLKL in Immunocytochemistry applications (Samson, A.L., et al. (2020). Nat Commun. 11(1):3151).
Immunofluorescence Analysis: A representative lot detected MLKL in Immunofluorescence applications (Samson, A.L., et al. (2020). Nat Commun. 11(1):3151).
Western Blotting Analysis: A representative lot detected MLKL in Western Blotting applications (Samson, A.L., et al. (2020). Nat Commun. 11(1):3151).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Anti-MLKL, clone 10C2, Cat. No. MABC1635, is a rat monoclonal antibody that detects MLKL and is tested for use in Immunocytochemistry, Immunofluorescence, and Western Blotting.
Purified rat monoclonal antibodyIgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Storage and Stability
Recommended storage: +2°C to +8°C.
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
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