Name: Anti-MLKL antibody [EPR17514]
Brand: RabMAb
SKU: ab184718
Rating: 5 (5 reviews)

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Western blot - Anti-MLKL antibody [EPR17514] (ab184718)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17514] to MLKL
Suitable for: ICC/IF, WB, IHC-P
Knockout validated
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

Product name

Anti-MLKL antibody [EPR17514]
See all MLKL primary antibodies
Description

Rabbit monoclonal [EPR17514] to MLKL
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, WB, IHC-Pmore details
Species reactivity

Reacts with: Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HUVEC, HT-29 and HeLa whole cell lysates; Human fetal kidney lysate. IHC-P: Human tonsil and colonic adenocarcinoma tissues.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17514
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human MLKL knockout HeLa cell line (ab255408)
KO cell lysates
- Human MLKL knockout HeLa cell lysate (ab263788)
KO cell pellets
- Human MLKL knockout HeLa cell pellet (ab278983)
Recombinant Protein
- Recombinant Human MLKL protein (ab153623)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab184718 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
ICC/IF	(1)	1/200.
WB	(4)	1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
IHC-P		1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Notes
ICC/IF 1/200.
WB 1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
IHC-P 1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Sequence similarities

Belongs to the protein kinase superfamily.
Contains 1 protein kinase domain.
Domain

The protein kinase domain is predicted to be catalytically inactive.
Target information above from: UniProt accession Q8NB16 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 197259 Human
- Omim: 615153 Human
- SwissProt: Q8NB16 Human
- Unigene: 119878 Human
Alternative names
- 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
see all

Western blot - Anti-MLKL antibody [EPR17514] (ab184718)

All lanes : Anti-MLKL antibody [EPR17514] (ab184718) at 1/1000 dilution

Lane 1 : HUVEC cell lysate
Lane 2 : HT-29 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : MLKL knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 54 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab184718 observed at 54 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab184718 was shown to react with MLKL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255408 (knockout cell lysate ab263788) was used. Wild-type and MLKL knockout samples were subjected to SDS-PAGE. ab184718 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-MLKL antibody [EPR17514] (ab184718)

All lanes : Anti-MLKL antibody [EPR17514] (ab184718) at 1/1000 dilution

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : MLKL knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Huvec cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 54 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab184718 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab184718 was shown to recognize MLKL in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when MLKL knockout samples were examined. Wild-type and MLKL knockout samples were subjected to SDS-PAGE. ab184718 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-MLKL antibody [EPR17514] (ab184718)

Ab184718 staining MLKL in HT-29 (Human colorectal adenocarcinoma epithelial cell) cells by Immunocytochemistry (ICC). Cells were fixed with 100% Methanol. Samples were incubated with primary antibody at 1/200 dilution (6.5μg/ml). An AlexaFluor^®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2μg/ml). Ab195889 , Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used as the counterstain antibody (1/200 dilution, 2.5 μg/ml . DAPI was used as a nuclear counterstain. Confocal image showing cytoplasmic staining on HT-29 cell line.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL antibody [EPR17514] (ab184718)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling MLKL with ab184718 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Cytoplasmic staining on the lymphocytes of human tonsil is observed. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL antibody [EPR17514] (ab184718)

Immunohistochemical analysis of paraffin-embedded Human colonic adenocarcinoma tissue labeling MLKL with ab184718 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Cytoplasmic staining on tumor cells of human colonic adenocarcinoma is observed. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-MLKL antibody [EPR17514] (ab184718)

All lanes : Anti-MLKL antibody [EPR17514] (ab184718) at 1/20000 dilution

Lane 1 : HUVEC (Human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : HT-29 (Human colorectal adenocarcinoma cells) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 54 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-MLKL antibody [EPR17514] (ab184718)

Anti-MLKL antibody [EPR17514] (ab184718) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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

Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-MLKL antibody [EPR17514] (ab184718)

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab184718? Please let us know so that we can cite the reference in this datasheet.

ab184718 has been referenced in 47 publications.

Liu B et al. Sodium iodate induces ferroptosis in human retinal pigment epithelium ARPE-19 cells. Cell Death Dis 12:230 (2021). PubMed: 33658488
Wang X et al. RIP3-mediated necroptosis was essential for spiral ganglion neuron damage. Neurosci Lett 744:135565 (2021). PubMed: 33359086
Rui C et al. The multitargeted kinase inhibitor KW-2449 ameliorates cisplatin-induced nephrotoxicity by targeting RIPK1-mediated necroptosis. Biochem Pharmacol 188:114542 (2021). PubMed: 33819469
Wang KJ et al. Emodin Induced Necroptosis and Inhibited Glycolysis in the Renal Cancer Cells by Enhancing ROS. Oxid Med Cell Longev 2021:8840590 (2021). PubMed: 33532038
Chen X et al. Heparanase induces necroptosis of microvascular endothelial cells to promote the metastasis of hepatocellular carcinoma. Cell Death Discov 7:33 (2021). PubMed: 33597510

View all Publications for this product

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1-5 of 5 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (HT-29)

Gel Running Conditions

Reduced Denaturing (8%)

Loading amount

20 µg

Treatment

1µM GNF-7 for 24hrs

Specification

HT-29

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

DR. Naghmeh Abiri

Verified customer

Submitted Mar 01 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (ovary)

Gel Running Conditions

Reduced Denaturing (12)

Loading amount

7 µg

Specification

ovary

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Konstantin Bagnjuk

Verified customer

Submitted Aug 04 2017

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa, HT29, L929, MEF)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

10 µg

Specification

HeLa, HT29, L929, MEF

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Xiaozheng Xu

Verified customer

Submitted Sep 08 2015

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HeLa)

Permeabilization

Yes - Triton X-100 0.25%

Specification

HeLa

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C

Fixative

Paraformaldehyde

Xiaozheng Xu

Verified customer

Submitted Sep 07 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa Cell)

Gel Running Conditions

Reduced Denaturing

Loading amount

20 µg

Specification

HeLa Cell

Abcam user community

Verified customer

Submitted Apr 24 2015

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

KO cell pellets

Recombinant Protein

Applications

The Abpromise guarantee

Target

Sequence similarities

Domain

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (47)

Customer reviews and Q&As

Western blot abreview for Anti-MLKL antibody [EPR17514]

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Western blot abreview for Anti-MLKL antibody [EPR17514]

Immunocytochemistry/ Immunofluorescence abreview for Anti-MLKL antibody [EPR17514]

Western blot abreview for Anti-MLKL antibody [EPR17514]