Overview

  • 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

  • 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.

Properties

  • 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

Applications

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.

Target

  • Sequence similarities
    Belongs to the protein kinase superfamily.
    Contains 1 protein kinase domain.
  • Domain
    The protein kinase domain is predicted to be catalytically inactive.
  • Information by UniProt
  • Database links
  • 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

Images

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

References

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

Customer reviews and Q&As

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1-4 of 4 Abreviews

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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