Mixed lineage kinase domain-like protein (UniProt: Q9D2Y4; also known as MLKL) is encoded by the Mlkl gene (Gene ID: 74568) in murine species. MLKL is a pseudokinase that plays a key role in TNF-induced necroptosis. MLKL is highly expressed in thymus, colon, intestine, liver, spleen and lung and is expressed at much lower level in skeletal muscle, heart and kidney. It is not detected in brain. MLKL is activated following phosphorylation by RIPK3 that induces a conformational switch, which is essential for necroptosis. The protein kinase domain of MLKL is catalytically inactive, but contains a pseudoactive site with an interaction between Lys-219 and Gln-343 residues. Upon phosphorylation by RIPK3, MLKL undergoes an active conformation and homotrimerization through its amino-terminal coiled-coil domain. Phosphorylation is also essential for homotrimerization of MLKL and its subsequent localization to the plasma membrane. Plasma membrane localization of MLKL is considered to be essential for Ca2+ influx, which is an early event of TNF-induced necroptosis. (Ref.: Cai, Z et al. (2014). Nat. Cell Biol. 16(1): 55-65).
Clone 7C6.1 specifically detects MLKL phosphorylalated on Ser345. It targets a sequence of 19 amino acids surrounding Ser345 in the C-terminal region.
KLH-conjugated linear peptide corresponding to 19 amino acids surrounding Ser345 from the C-terminal half of murine Mixed lineage kinase domain-like protein (MLKL).
ELISA Analysis: A representative lot detected phospho-MLKL (Ser345) in ELISA applications (Rodriguez, D.A., et. al. (2016). 23(1):76-88).
Immunocytochemistry Analysis: A representative lot detected phospho-MLKL (Ser345) in Immunocytochemistry applications (Rodriguez, D.A., et. al. (2016). 23(1):76-88).
Western Blotting Analysis: A representative lot detected phospho-MLKL (Ser345) in Western Blotting applications (Rodriguez, D.A., et. al. (2016). 23(1):76-88).
Detect phospho MLKL Ser345 using this mouse monoclonal Anti-phospho-MLKL (Ser345), clone 7C6.1, Cat. No. MABC1158. It has been tested in ELISA, Immunocytochemistry, and Western Blotting.
Apoptosis & Cancer
Evaluated by Western Blotting in MLKL -/- mouse embryonic fibroblasts expressing doxycycline-inducible wild-type MLKL and treated with TNF alpha and Z-VAD-FMK.
Western Blotting Analysis: A 1:4,000 dilution of this antibody detected phospho-MLKL Ser345 in 20 ug of lysate from MLKL -/- mouse embryonic fibroblasts expressing doxycycline-inducible wild-type MLKL-Flag-C construct and treated with TNF alpha (10 ng/mL) and Z-VAD-FMK (25 uM).
~54 kDa observed; 54.32 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
Protein G purified
Purified mouse monoclonal antibody IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Storage and Stability
Stable for 1 year at 2-8°C from date of receipt.
Concentration: Please refer to lot specific datasheet.
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