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Western blot(SDS PAGE) analysis of extracts fromHT-29 cells, treated with Z-VAD (20 μM, 30 min )、hTNF-α(20 ng/ml, 7 hr)、SM-164 (100 nM, 7 hr).Using Phospho-MLKL (Ser358) Rabbit mAb IgG [SSM7] at dilution of 1:1000 incubated at 4℃ over night.
Protein names :MLKL,MLKL_HUMAN,Mixed lineage kinase domain-like protein
UniProtID :Q8NB16
MASS(da) :54,479
MW(kDa) : 54
Form :Liquid
Purification :Protein A purification
Host :Rabbit
Isotype :IgG
sensitivity :Endogenous
Reactivity :Human
Specificity :Antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser358 of human MLKL
Storage :Antibody store in 10 mM PBS, 0.5mg/ml BSA, 50% glycerol. Shipped at 4°C. Store at-20°C or -80°C. Products are valid for one natural year of receipt.Avoid repeated freeze / thaw cycles.
WB Positive detected : HT-29 cells, treated with Z-VAD (20 μM, 30 min )、hTNF-α(20 ng/ml, 7 hr)、SM-164 (100 nM, 7 hr)
Function : Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process (PubMed:22265413, PubMed:22265414, PubMed:22421439, PubMed:24316671). Does not have protein kinase activity (PubMed:22265413, PubMed:22265414, PubMed:22421439, PubMed:24316671). Activated following phosphorylation by RIPK3, leading to homotrimerization, localization to the plasma membrane and execution of programmed necrosis characterized by calcium influx and plasma membrane damage (PubMed:22265413, PubMed:22265414, PubMed:22421439, PubMed:24316671). In addition to TNF-induced necroptosis, necroptosis can also take place in the nucleus in response to orthomyxoviruses infection: following activation by ZBP1, MLKL is phosphorylated by RIPK3 in the nucleus, triggering disruption of the nuclear envelope and leakage of cellular DNA into the cytosol.following ZBP1 activation, which senses double-stranded Z-RNA structures, nuclear RIPK3 catalyzes phosphorylation and activation of MLKL, promoting disruption of the nuclear envelope and leakage of cellular DNA into the cytosol (By similarity). Binds to highly phosphorylated inositol phosphates such as inositolhexakisphosphate (InsP6) which is essential for its necroptotic function (PubMed:29883610)..
Subcellular locationi :Cytoplasm. Cell membrane. Nucleus.
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 1% w/v BSA, 1X TBST at 4°C overnight.