MLKL Monoclonal antibody

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  WB analysis using 66675-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66675-1-Ig (MLKL antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 66675-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66675-1-Ig (MLKL antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

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  WB analysis of 4T1 using 66675-1-Ig

  4T1 cells were subjected to SDS PAGE followed by western blot with 66675-1-Ig (MLKL antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IHC staining of human liver cancer using 66675-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66675-1-Ig (MLKL antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 66675-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66675-1-Ig (MLKL antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 66675-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66675-1-Ig (MLKL antibody) at dilution of 1:300 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66675-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66675-1-Ig (MLKL antibody) at dilution of 1:300 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human colon cancer using 66675-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 66675-1-Ig (MLKL antibody) at dilution of 1:300 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human colon cancer using 66675-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 66675-1-Ig (MLKL antibody) at dilution of 1:300 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of human liver cancer using 66675-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human liver cancer tissue using 66675-1-Ig (MLKL antibody) at dilution of 1:100 and CoraLite488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  IF Staining of HepG2 using 66675-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using MLKL antibody (66675-1-Ig, Clone: 3D4C6 ) at dilution of 1:800 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (RGAM002).

• Featured Product
• KD/KO Validated

MLKL Monoclonal Antibody for WB, IHC, IF/ICC, IF-P, ELISA

Cat No. 66675-1-Ig

Clone No.3D4C6

Publications(125)

Reviews (2)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse and More (2)

Applications

WB, IHC, IF/ICC, IF-P, IP, CoIP, ELISA

Synonyms

3D4C6, hMLKL, Mixed lineage kinase domain-like protein

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Tested Applications

• Positive WB detected in: HT-29 cells, 4T1 cells, PC-3 cells, HeLa cells, HepG2 cells, NIH/3T3 cells, mMSCs cells
• Positive IHC detected in: human liver cancer tissue, human colon cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human liver cancer tissue
• Positive IF/ICC detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 4 publications below
• WB: See 112 publications below
• IHC: See 13 publications below
• IF: See 18 publications below
• IP: See 2 publications below
• CoIP: See 2 publications below

Product Information

66675-1-Ig targets MLKL in WB, IHC, IF/ICC, IF-P, IP, CoIP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, mouse, rat, zebrafish
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag15188

  Product name: Recombinant human MLKL protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 1-263 aa of BC028141

  Sequence: MENLKHIITLGQVIHKRCEEMKYCKKQCRRLGHRVLGLIKPLEMLQDQGKRSVPSEKLTTAMNRFKAALEEANGEIEKFSNRSNICRFLTASQDKILFKDVNRKLSDVWKELSLLLQVEQRMPVSPISQGASWAQEDQQDADEDRRAFQMLRRDNEKIEASLRRLEINMKEIKETLRQSLESSSGKSPLEISRFKVKNVKTGSASGCNSEKIRKLVAVKRQQEPLGEDCPSELREIIDECRAHDPSVRPSVDEILKKLSTFSK

• Full Name: mixed lineage kinase domain-like
• Calculated Molecular Weight: 471 aa, 54 kDa
• Observed Molecular Weight: 35-40 kDa, 50-55 kDa
• GenBank Accession Number: BC028141
• Gene Symbol: MLKL
• Gene ID (NCBI): 197259
• RRID: AB_2882029
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q8NB16
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Mixed lineage kinase domain like pseudokinase (MLKL), belongs to the protein kinase superfamily and has two MW of 54 and 30 kDa. MLKL 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 the necroptosis pathway. High levels of this protein and RIP3 are associated with inflammatory bowel disease in children. The 66675-1-Ig antibody recognizes 54 kDa MLKL monomer, 216 kDa MLKL tetramer, and a band around 45-50 kDa which are similar to papers published. (PMID: 31848291). MLKL can be detected as a 35kDa truncated isoform (PMID: 35965541).

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	IF	Adv Sci (Weinh); Pressure Drives Rapid Burst-Like Coordinated Cellular Motion from 3D Cancer Aggregates.; Authors - Swetha Raghuraman
mouse,human	WB	Acta Pharm Sin B; Ligand-based substituent-anchoring design of selective receptor-interacting protein kinase 1 necroptosis inhibitors for ulcerative colitis therapy; Authors - Jing Zhu
mouse	WB,IHC	Acta Pharm Sin B; A new perspective of triptolide-associated hepatotoxicity: the relevance of NF- κ B and NF- κ B-mediated cellular FLICE-inhibitory protein.; Authors - Ziqiao Yuan
human	WB	Aging Dis; MiR-29a-3p Improves Acute Lung Injury by Reducing Alveolar Epithelial Cell PANoptosis.; Authors - Yanhui Cui
mouse,human	WB	Cell Death Dis; S100A8/A9hi neutrophils induce mitochondrial dysfunction and PANoptosis in endothelial cells via mitochondrial complex I deficiency during sepsis; Authors - Yanghanzhao Wang
mouse	WB	Free Radic Biol Med; Short-term exposure to low doses of aflatoxin B1 aggravates nonalcoholic steatohepatitis by TLR4-mediated necroptosis; Authors - Junya Zeng

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Fan (Verified Customer) (03-22-2021)

Mouse retinal tissue was used. Multiple bands were detected. The target band has the strongest intensity.

• Applications: Western Blot
• Primary Antibody Dilution: 1:500

MLKL Antibody Western Blot validation (1:500 dilution) in (Cat no:66675-1-Ig)

FH

Paramananda (Verified Customer) (01-31-2019)

Excellent antibody!

• Applications: Immunohistochemistry,
• Primary Antibody Dilution: 1:50
• Cell Tissue Type: Cornea