MLKL Antibody - #DF7412

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Product: MLKL Antibody
Catalog: DF7412
Description: Rabbit polyclonal antibody to MLKL
Application: WB IHC IF/ICC
Cited expt.: WB, IHC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 54kDa; 54kD(Calculated).
Uniprot: Q8NB16
RRID: AB_2839350

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MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
MLKL Antibody detects endogenous levels of total MLKL.
RRID:
AB_2839350
Cite Format: Affinity Biosciences Cat# DF7412, RRID:AB_2839350.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

9130019I15Rik; FLJ34389; hMLKL; Mixed lineage kinase domain like; Mixed lineage kinase domain like protein; Mixed lineage kinase domain like pseudokinase; Mixed lineage kinase domain-like protein; Mlkl; MLKL_HUMAN;

Immunogens

Immunogen:

A synthesized peptide derived from human MLKL, corresponding to a region within the internal amino acids.

Uniprot:

Q8NB16(MLKL_HUMAN) >>Visit HPA database.

Gene(ID):
MLKL(197259)
Sequence:
MENLKHIITLGQVIHKRCEEMKYCKKQCRRLGHRVLGLIKPLEMLQDQGKRSVPSEKLTTAMNRFKAALEEANGEIEKFSNRSNICRFLTASQDKILFKDVNRKLSDVWKELSLLLQVEQRMPVSPISQGASWAQEDQQDADEDRRAFQMLRRDNEKIEASLRRLEINMKEIKETLRQYLPPKCMQEIPQEQIKEIKKEQLSGSPWILLRENEVSTLYKGEYHRAPVAIKVFKKLQAGSIAIVRQTFNKEIKTMKKFESPNILRIFGICIDETVTPPQFSIVMEYCELGTLRELLDREKDLTLGKRMVLVLGAARGLYRLHHSEAPELHGKIRSSNFLVTQGYQVKLAGFELRKTQTSMSLGTTREKTDRVKSTAYLSPQELEDVFYQYDVKSEIYSFGIVLWEIATGDIPFQGCNSEKIRKLVAVKRQQEPLGEDCPSELREIIDECRAHDPSVRPSVDEILKKLSTFSK

Research Backgrounds

Function:

Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process. 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. Does not have protein kinase activity. Binds to highly phosphorylated inositol phosphates such as inositolhexakisphosphate (InsP6) which is essential for its necroptotic function.

PTMs:

Phosphorylation by RIPK3 induces a conformational switch that is required for necroptosis. It also induces homotrimerization and localization to the plasma membrane.

Subcellular Location:

Cytoplasm. Cell membrane.
Note: Localizes to the cytoplasm and translocates to the plasma membrane on necroptosis induction.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Family&Domains:

The protein kinase domain is catalytically inactive but contains an unusual pseudoactive site with an interaction between Lys-230 and Gln-356 residues. Upon phosphorylation by RIPK3, undergoes an active conformation (By similarity).

The coiled coil region 2 is responsible for homotrimerization.

Belongs to the protein kinase superfamily.

Research Fields

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway.   (View pathway)

References

1). Cyclic helix B peptide alleviates proinflammatory cell death and improves functional recovery after traumatic spinal cord injury. Redox Biology, 2023 (PubMed: 37290302) [IF=10.7]
2). Fetuin-A alleviates neuroinflammation against traumatic brain injury-induced microglial necroptosis by regulating Nrf-2/HO-1 pathway. Journal of neuroinflammation, 2022 (PubMed: 36333786) [IF=9.3]
3). Circular RNA FEACR inhibits ferroptosis and alleviates myocardial ischemia/reperfusion injury by interacting with NAMPT. Journal of biomedical science, 2023 (PubMed: 37370086) [IF=9.0]
4). Melatonin restores endoplasmic reticulum homeostasis to protect injured neurons in a rat model of chronic cervical cord compression. JOURNAL OF PINEAL RESEARCH, 2023 (PubMed: 36732085) [IF=8.3]
5). Glycidol induces necroptosis and inflammation through autophagy-necrosome pathway in renal cell and mice. The Science of the total environment, 2025 (PubMed: 39965374) [IF=8.2]
6). Target deubiquitinase OTUB1 as a therapeatic strategy for BLCA via β-catenin/necroptosis signal pathway. International journal of biological sciences, 2024 (PubMed: 39113709) [IF=8.2]

Application: WB    Species: Mouse    Sample: BLCA cells

Figure 7. OTUB1 is involved in cisplatin resistance of BLCA through β-catenin stabilization. A. Cell survival assay was determined in T24 control cell and T24 cisplatin-resistance cell. B. The changes in cell morphology between the T24 control cell and the T24 cisplatin-resistance cell. C. Relative expression of several chemoresistance markers (including MDR1, BCRP and YB-1) in T24 control cell and T24 cisplatin-resistance cell with or without cisplatin treatment. D. Relative expression of OTUB1 in T24 control cell and T24 cisplatin-resistance cell with or without cisplatin treatment. E. Relative expression of OTUB1, necroptosis-related markers (such as RIPK3, MLKL and P-MLKL), β-catenin and downstream proteins (including AXIN-2, C-myc, cyclin D1 and TCF1) in T24 cisplatin-resistance cells treated with gradient cisplatin concentration. F. Immunoprecipitation assay showed the relationship between OTUB1 and β-catenin in T24 cisplatin-resistance cells. G. Immunoprecipitation assay showed that gradient cisplatin concentration promotes the interaction between OTUB1 and β-catenin, and restrains the ubiquitination of β-catenin. H. Relative expression of OTUB1, necroptosis-related markers (such as RIPK3, MLKL and P-MLKL), β-catenin and downstream targets following elevated OTUB1, β-catenin with or without XAV-939 treatment in T24 cisplatin-resistance cell (control, OTUB1, OTUB1/XAV-939, β-catenin). I. Cell survival assay was determined in T24 cisplatin-resistance cells following overexpressed OTUB1 with or without 10uM XAV-939 (or knockdown OTUB1 with or without overexpressed β-catenin). J. Knockdown OTUB1 restrains the growth in T24 cisplatin-resistance mice bladder tumor in vivo.
7). NAD+-boosting agent nicotinamide mononucleotide potently improves mitochondria stress response in Alzheimer's disease via ATF4-dependent mitochondrial UPR. Cell death & disease, 2024 (PubMed: 39394148) [IF=8.1]
8). Proanthocyanidins alleviate acute alcohol liver injury by inhibiting pyroptosis via inhibiting the ROS-MLKL-CTSB-NLRP3 pathway. Phytomedicine : international journal of phytotherapy and phytopharmacology, 2025 (PubMed: 39612889) [IF=7.9]
9). Polystyrene nanoplastics and cadmium co-exposure aggravated cardiomyocyte damage in mice by regulating PANoptosis pathway. Environmental pollution (Barking, Essex : 1987), 2024 (PubMed: 38462200) [IF=7.6]
10). Activation of ALDH2 attenuates high glucose induced rat cardiomyocyte fibrosis and necroptosis. Free radical biology & medicine, 2020 (PubMed: 31689484) [IF=7.1]

Application: WB    Species: rat    Sample: cardiomyocytes

Fig. 8 |Changes of RIP1, RIP3 and MLKL at mRNA and protein levels in the different groups (means ± SD, n=6) A, B, C: RIP1, RIP3 and MLKL mRNA levels in cardiomyocytes normalized by GAPDH levels. **P<0.01 compared with NG; #P<0.05, ##P<0.01 compared with HG; ^P<0.05, ^^P<0.01 compared with HG+Alda-1 D: Representative western blots of RIP1, RIP3 and MLKL proteins
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