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  1. Link

    mlkl-antibody-epr17514-ab184718.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases Other Kinases
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MLKL antibody [EPR17514] (ab184718)

  • Datasheet
  • Certificate of Compliance
Reviews (5) Submit a question References (47)

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Western blot - Anti-MLKL antibody [EPR17514] (ab184718)
  • Western blot - Anti-MLKL antibody [EPR17514] (ab184718)
  • Immunocytochemistry/ Immunofluorescence - Anti-MLKL antibody [EPR17514] (ab184718)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL antibody [EPR17514] (ab184718)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL antibody [EPR17514] (ab184718)
  • Western blot - Anti-MLKL antibody [EPR17514] (ab184718)
  • Western blot - Anti-MLKL antibody [EPR17514] (ab184718)
  • 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

Conjugates logo Related conjugates and formulations

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

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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. 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.

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

    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

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Cell Death
    • Necroptosis

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-MLKL antibody [EPR17514] (ab207901)
    • Alexa Fluor® 647 Anti-MLKL antibody [EPR17514] (ab207902)
    • Alexa Fluor® 594 Anti-MLKL antibody [EPR17514] (ab208082)
    • Anti-MLKL antibody [EPR17514] - BSA and Azide free (ab211045)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • 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)

Applications

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.

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.
  • 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

    Images

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

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (47)

    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

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-5 of 5 Abreviews or Q&A

    Western blot abreview for Anti-MLKL antibody [EPR17514]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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%
    Read More

    DR. Naghmeh Abiri

    Verified customer

    Submitted Mar 01 2021

    Western blot abreview for Anti-MLKL antibody [EPR17514]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Konstantin Bagnjuk

    Verified customer

    Submitted Aug 04 2017

    Western blot abreview for Anti-MLKL antibody [EPR17514]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Xiaozheng Xu

    Verified customer

    Submitted Sep 08 2015

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Xiaozheng Xu

    Verified customer

    Submitted Sep 07 2015

    Western blot abreview for Anti-MLKL antibody [EPR17514]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa Cell)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    HeLa Cell
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 24 2015

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