Name: Anti-MLKL antibody [EPR17514]
Brand: RabMAb
SKU: ab184718
Rating: 5 (4 reviews)

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 within Human MLKL aa 350 to the C-terminus. The exact sequence is proprietary.
Database link: Q8NB16

Run BLAST with Run BLAST with
Positive control
- WB: HUVEC and HT-29 whole cell lysates; Human fetal kidney lysate. IHC-P: Human tonsil and colonic adenocarcinoma tissues.
General notes

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.
This product is a recombinant rabbit monoclonal antibody.

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

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

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		1/200.
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)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MLKL knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Huvec cell lysate (20 µg)
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.
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.
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.

Western blot protocols
Immunohistochemistry protocols

Datasheet

This product has been referenced in:

Saito Y et al. Amino acid starvation culture condition sensitizes EGFR-expressing cancer cell lines to gefitinib-mediated cytotoxicity by inducing atypical necroptosis. Int J Oncol 52:1165-1177 (2018). Read more (PubMed: 29484439) »
Xu Q et al. The cytoplasmic nuclear receptor RAR? controls RIP1 initiated cell death when cIAP activity is inhibited. Nat Commun 8:425 (2017). Read more (PubMed: 28871172) »

See all 3 Publications for this product

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

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Abcam guarantees this product to work in the species/application used in this Abreview.

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 AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-MLKL antibody [EPR17514] (ab184718)

Overview

Properties

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Protocols

Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

Western blot abreview for Anti-MLKL antibody [EPR17514]

Western blot abreview for Anti-MLKL antibody [EPR17514]

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

Western blot abreview for Anti-MLKL antibody [EPR17514]