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Phospho-MLKL (S345) Recombinant Rabbit Monoclonal Antibody [JM92-37]

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Specification

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Catalog# ET1705-51

Phospho-MLKL (S345) Recombinant Rabbit Monoclonal Antibody [JM92-37]

Application

  • WB

  • IHC-Fr

Reactivity

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-MLKL (S345) Recombinant Rabbit Monoclonal Antibody [JM92-37]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser345 of mouse MLKL.

Species Reactivity

Mouse

Validated Applications

WB, IHC-Fr

Molecular Weight

Predicted band size: 54 kDa

Positive Control

L929 treated with 20μM Z-VAD for 3.5 hours then added 100nM SM-164 and 20ng/ml TNF-α for 3 hours cell lysate.

Conjugation

unconjugated

Clone Number

JM92-37

RRID

AB_3070579

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-Fr

  • 1:100

Target

Function

Mixed lineage kinase domain like pseudokinase (MLKL) is a protein that in humans is encoded by the MLKL gene. This gene belongs to the protein kinase superfamily. The encoded protein contains a protein kinase-like domain; however, is thought to be inactive because it lacks several residues required for activity. This protein plays a critical role in tumor necrosis factor (TNF)-induced necroptosis, a programmed cell death process, via interaction with receptor-interacting protein 3 (RIP3), which is a key signaling molecule in necroptosis pathway. Inhibitor studies and knockdown of this gene inhibited TNF-induced necrosis. High levels of this protein and RIP3 are associated with inflammatory bowel disease in children.

Background References

1. Martens S et al. MLKL in cancer: more than a necroptosis regulator. Cell Death Differ. 2021 Jun

2. Liu S et al. MLKL polymerization-induced lysosomal membrane permeabilization promotes necroptosis. Cell Death Differ. 2024 Jan

Sequence Similarity

Belongs to the protein kinase superfamily.

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.

Post-translational Modification

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

Subcellular Location

Cell membrane, Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q9D2Y4 Mouse

Synonyms

9130019I15Rik antibody

FLJ34389 antibody

hMLKL antibody

Mixed lineage kinase domain like antibody

Mixed lineage kinase domain like protein antibody

Mixed lineage kinase domain like pseudokinase antibody

Mixed lineage kinase domain-like protein antibody

Mlkl antibody

MLKL_HUMAN antibody

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-MLKL (S345) on different lysates with Rabbit anti-Phospho-MLKL (S345) antibody (<a href="/products/ET1705-51" style="font-weight: bold;text-decoration: underline;">ET1705-51</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: L-929 cell lysate<br />Lane 2: L929 treated with 20μM Z-VAD for 3.5 hours, then added 100nM SM-164 and 20ng/ml TNF-α for 3 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 54 kDa<br /><br />Exposure time: 1 minute 10 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1705-51" style="font-weight: bold;text-decoration: underline;">ET1705-51</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-MLKL (S345) on different lysates with Rabbit anti-Phospho-MLKL (S345) antibody (ET1705-51) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: L-929 cell lysate
    Lane 2: L929 treated with 20μM Z-VAD for 3.5 hours, then added 100nM SM-164 and 20ng/ml TNF-α for 3 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 54 kDa

    Exposure time: 1 minute 10 seconds; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-51) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-Phospho-MLKL (S345) antibody (<a href="/products/ET1705-51" style="font-weight: bold;text-decoration: underline;">ET1705-51</a>) at 1/100 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ET1705-51" style="font-weight: bold;text-decoration: underline;">ET1705-51</a>, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-Phospho-MLKL (S345) antibody (ET1705-51) at 1/100 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1705-51, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation