Phospho-MLKL (Ser345) Antibody - #AF3902

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Product: Phospho-MLKL (Ser345) Antibody
Catalog: AF3902
Description: Rabbit polyclonal antibody to Phospho-MLKL (Ser345)
Application: ELISA(peptide)
Cited expt.:
Reactivity: Mouse
Mol.Wt.: 54kD(Calculated).
Uniprot: Q9D2Y4
RRID: AB_2847216

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

Product Info

Source:
Rabbit
Application:
ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*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:
Mouse
Clonality:
Polyclonal
Specificity:
Phospho-MLKL (Ser345) Antibody detects endogenous levels of MLKL only when phosphorylated at Ser345.
RRID:
AB_2847216
Cite Format: Affinity Biosciences Cat# AF3902, RRID:AB_2847216.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
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.

Immunogens

Immunogen:

A synthesized peptide derived from mouse MLKL around the phosphorylation site of Ser345.

Uniprot:

Q9D2Y4(MLKL_MOUSE) >>Visit HPA database.

Expression:
Q9D2Y4 MLKL_MOUSE:

Highly expressed in thymus, colon, intestine, liver, spleen and lung. Expressed at much lower level in skeletal muscle, heart and kidney. Not detected in brain.

Sequence:
MDKLGQIIKLGQLIYEQCEKMKYCRKQCQRLGNRVHGLLQPLQRLQAQGKKNLPDDITAALGRFDEVLKEANQQIEKFSKKSHIWKFVSVGNDKILFHEVNEKLRDVWEELLLLLQVYHWNTVSDVSQPASWQQEDRQDAEEDGNENMKVILMQLQISVEEINKTLKQCSLKPTQEIPQDLQIKEIPKEHLGPPWTKLKTSKMSTIYRGEYHRSPVTIKVFNNPQAESVGIVRFTFNDEIKTMKKFDSPNILRIFGICIDQTVKPPEFSIVMEYCELGTLRELLDREKDLTMSVRSLLVLRAARGLYRLHHSETLHRNISSSSFLVAGGYQVKLAGFELSKTQNSISRTAKSTKAERSSSTIYVSPERLKNPFCLYDIKAEIYSFGIVLWEIATGKIPFEGCDSKKIRELVAEDKKQEPVGQDCPELLREIINECRAHEPSQRPSVDGRSLSGRERILERLSAVEESTDKKV

Research Backgrounds

Function:

Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process. Does not have protein kinase activity. 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. In addition to TNF-induced necroptosis, necroptosis can also take place in the nucleus in response to orthomyxoviruses infection: 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. Binds to highly phosphorylated inositol phosphates such as inositolhexakisphosphate (InsP6) which is essential for its necroptotic function (By similarity).

PTMs:

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

Subcellular Location:

Cytoplasm. Cell membrane. Nucleus.
Note: Localizes to the cytoplasm and translocates to the plasma membrane on necroptosis induction (By similarity). Localizes to the nucleus in response to orthomyxoviruses infection (PubMed:32200799).

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

Highly expressed in thymus, colon, intestine, liver, spleen and lung. Expressed at much lower level in skeletal muscle, heart and kidney. Not detected in brain.

Family&Domains:

The coiled coil region 2 is responsible for homotrimerization.

The protein kinase domain is catalytically inactive but contains an unusual pseudoactive site with an interaction between Lys-219 and Gln-343 residues (PubMed:24012422, PubMed:24095729). Upon phosphorylation by RIPK3, undergoes an active conformation (PubMed:24012422, PubMed:24095729).

Belongs to the protein kinase superfamily.

References

1). Apoptotic Bodies Derived from Fibroblast-Like Cells in Subcutaneous Connective Tissue Inhibit Ferroptosis in Ischaemic Flaps via the miR-339-5p/KEAP1/Nrf2 Axis. Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2024 (PubMed: 38639443) [IF=15.1]
2). Inhibition of PLA2G4E/cPLA2 promotes survival of random skin flaps by alleviating Lysosomal membrane permeabilization-Induced necroptosis. Autophagy, 2022 (PubMed: 34872436) [IF=14.6]

Application: IF/ICC    Species: human    Sample: HUVECs

Figure 8. Overexpression of Mir504-5p suppressed PLA2G4E and protected the function of HUVECs. (A) Immunofluorescence staining of LAMP1 and p-PLA2G4E in HUVECs exposed to different treatments for 24 h (NC, TBHP, TBHP + Mir-NC or TBHP + Mir504-5p mimic). Scale bars: 20 μm. (B) Comparison of the number of p-PLA2G4E-positive lysosomes in each HUVEC among the four groups. Data are expressed as the means ± SEM (n = 6). (C) Immunofluorescence staining of LAMP1 and CTSD in HUVECs exposed to different treatments for 24 h (NC, TBHP, TBHP + Mir-NC or TBHP + Mir504-5p mimic). Scale bars: 20 μm. (D) Comparison of the ratio of diffuse CTSD cells (refer to HUVECs) among the four groups. Data are expressed as the means ± SEM (n = 6). (E) Immunofluorescence staining of p-MLKL in HUVECs exposed to different treatments for 24 h (NC, TBHP, TBHP + Mir-NC or TBHP + Mir504-5p mimic). Scale bars: 20 μm. (F) Quantification of the integrated intensity of p-MLKL in HUVECs. Data are expressed as the means ± SEM (n = 6). (G) Cell migration assays were performed on HUVECs after 24 h of different treatments (NC, TBHP, TBHP + Mir-NC or TBHP + Mir504-5p mimic), and the presented results were obtained after 12 h of culture. Scale bars: 200 μm. (H) Quantification and analysis of the number of migrated cells (refer to HUVECs). Data are expressed as the means ± SEM (n = 6). (I) An in vitro angiogenesis (tube formation) assay was performed on HUVECs after 24 h of different treatments (NC, TBHP, TBHP + Mir-NC or TBHP + Mir504-5p mimic), and the presented results were obtained after 6 h of culture. Scale bars: 200 μm. (J) Quantification and analysis of tube length (pixels; ×104). Data are expressed as the means ± SEM (n = 6). Significance: *p < 0.05.
3). The critical role of necroptosis in oil cyst formation associated with autologous fat transplantation. Cell communication and signaling : CCS, 2026 (PubMed: 41857749) [IF=8.4]
4). MLKL signaling regulates macrophage polarization in acute pancreatitis through CXCL10. Cell death & disease, 2023 (PubMed: 36828808) [IF=8.1]

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