Skip to main content

Anti-MLKL antibody is a rabbit monoclonal antibody that is used in MLKL western blot (WB), immunofluorescence (ICC/IF), and IHC. Suitable for human samples.


- Cited in over 80 publications
- Affinity-purified using Protein A

Images

Western blot - Anti-MLKL antibody [EPR17514] (AB184718), expandable thumbnail
  • Western blot - Anti-MLKL antibody [EPR17514] (AB184718), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MLKL antibody [EPR17514] (AB184718), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL antibody [EPR17514] (AB184718), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL antibody [EPR17514] (AB184718), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFIHC-P
Human
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info
1/1000
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info
1/200
Notes

-

Tested
Tested

Species

Human

Dilution info
1/400
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

9 products for Alternative Product

Target data

Function

Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process (PubMed:22265413, PubMed:22265414, PubMed:22421439, PubMed:24316671). Does not have protein kinase activity (PubMed:22265413, PubMed:22265414, PubMed:22421439, PubMed:24316671). Activated following phosphorylation by RIPK3, leading to homotrimerization, localization to the plasma membrane and execution of programmed necrosis characterized by calcium influx and plasma membrane damage (PubMed:22265413, PubMed:22265414, PubMed:22421439, PubMed:24316671). In addition to TNF-induced necroptosis, necroptosis can also take place in the nucleus in response to orthomyxoviruses infection: following activation by ZBP1, MLKL is phosphorylated by RIPK3 in the nucleus, triggering disruption of the nuclear envelope and leakage of cellular DNA into the cytosol.following ZBP1 activation, which senses double-stranded Z-RNA structures, nuclear RIPK3 catalyzes phosphorylation and activation of MLKL, promoting disruption of the nuclear envelope and leakage of cellular DNA into the cytosol (By similarity). Binds to highly phosphorylated inositol phosphates such as inositolhexakisphosphate (InsP6) which is essential for its necroptotic function (PubMed:29883610).

Alternative names

Mixed lineage kinase domain-like protein, hMLKL, MLKL

Recommended products

Anti-MLKL antibody is a rabbit monoclonal antibody that is used in MLKL western blot (WB), immunofluorescence (ICC/IF), and IHC. Suitable for human samples.


- Cited in over 80 publications
- Affinity-purified using Protein A

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR17514

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Product Specifications

Anti-MLKL antibody [EPR17514] (ab184718) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF, IHC-P, WB in human samples.
Anti-MLKL antibody [EPR17514] (ab184718) specifically detects MLKL (UniProt ID: Q8NB16; Molecular weight: 54kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation

Abcam's high quality manufacturing and validation processes ensure Anti-MLKL antibody [EPR17514] (ab184718) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-MLKL antibody [EPR17514] (ab184718) has been confirmed by testing in knockout samples.
Anti-MLKL antibody [EPR17514] (ab184718) has been cited over 84 times in peer reviewed journals and is trusted by the scientific community.
Anti-MLKL antibody [EPR17514] (ab184718) has 5 independent reviews from customers.
Related Products
Conjugation-ready, carrier free format available for antibody clone EPR17514 - Anti-MLKL antibody [EPR17514] - BSA and Azide free ab211045.
Antibody clone EPR17514 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 594 (Alexa Fluor® 488 Anti-MLKL antibody [EPR17514] ab207901, Alexa Fluor® 647 Anti-MLKL antibody [EPR17514] ab207902, Alexa Fluor® 594 Anti-MLKL antibody [EPR17514] ab208082).

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MLKL also known as mixed lineage kinase domain-like protein plays a critical role in the process of necroptosis a form of programmed cell death. The MLKL protein has a molecular weight of approximately 54 kDa. The protein exists mainly within the cytoplasm but translocates to the plasma membrane during cell death execution. Expression of MLKL happens in various tissues indicating its wide biological importance. Phosphorylation of MLKL often referred to as p-MLKL is key to triggering its activity marking the transition from an inactive to an active state during necroptosis.

Biological function summary

The MLKL protein acts as an executioner of cell death by forming a complex that disrupts the plasma membrane integrity. This process is downstream of receptor-interacting serine/threonine-protein kinase 3 (RIPK3) which phosphorylates MLKL to form the active necrosome complex. Active MLKL oligomerizes and migrates towards the inner leaflet of the plasma membrane binding to phosphatidylinositol phosphates which assists in pore formation and cellular rupture. The ability to measure MLKL activity levels such as via MLKL ELISA kits is important for understanding necrotic processes in detailed studies.

Pathways

MLKL is integrally involved in the necroptotic pathway alongside RIPK1 and RIPK3 which are key initiators of necroptosis. Phosphorylated MLKL acts downstream of RIPK3 resulting in cell death without caspase activation distinguishing necroptosis from apoptosis. MLKL and RIPK3 are tightly linked within this pathway with MLKL phosphorylation serving as a vital event for the execution phase. The necroptosis pathway is part of larger networks including inflammatory response pathways highlighting the importance of MLKL's role beyond sheer cell death.

Associated diseases and disorders

MLKL has been implicated in various inflammatory conditions and neurodegenerative diseases. The dysregulation of necroptosis can contribute to disorders such as inflammatory bowel disease and amyotrophic lateral sclerosis. In inflammatory bowel disease increased levels of p-MLKL might lead to excessive cell death exacerbating inflammation. Similarly in neurodegenerative disorders the harmful activation of MLKL may accelerate neuronal cell death. Key interactions with proteins like RIPK3 and RIPK1 highlight MLKL's involvement in these pathological processes making it a potential target for therapeutic intervention.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

7 product images

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

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

    Lanes 1 - 4: Merged signal (red and green). Green - ab184718 observed at 54 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Human MLKL knockout HeLa cell line ab255408 (knockout cell lysate Human MLKL knockout HeLa cell lysate ab263788) was used. Wild-type and MLKL knockout samples were subjected to SDS-PAGE. ab184718 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

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

    Lane 1: HUVEC cell lysate at 20 µg

    Lane 2: HT-29 cell lysate at 20 µg

    Lane 2: Western blot - Human MLKL knockout HeLa cell line (Human MLKL knockout HeLa cell line ab255408)

    Lane 3: Wild-type HeLa cell lysate at 20 µg

    Lane 4: MLKL knockout HeLa cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 54 kDa

    Observed band size: 37 kDa, 54 kDa

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

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

    Lanes 1 - 4: Merged signal (red and green). Green - ab184718 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

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

    Lane 1: Wild-type HAP1 cell lysate at 20 µg

    Lane 2: MLKL knockout HAP1 cell lysate at 20 µg

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: Huvec cell lysate at 20 µg

    Predicted band size: 54 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-MLKL antibody [EPR17514] (ab184718), expandable thumbnail

    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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 dilution (2μg/ml). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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), expandable thumbnail

    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 (Goat Anti-Rabbit IgG H&L (HRP) 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) (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), expandable thumbnail

    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 (Goat Anti-Rabbit IgG H&L (HRP) 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) (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), expandable thumbnail

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

    Blocking/Dilution buffer: 5% NFDM/TBST.

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

    Lane 1: HUVEC (Human umbilical vein endothelial cell line) whole cell lysate at 20 µg

    Lane 2: HT-29 (Human colorectal adenocarcinoma cells) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 54 kDa

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

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

    Blocking/Dilution buffer: 5% NFDM/TBST.

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

    All lanes: Human fetal kidney lysate at 10 µg

    Secondary

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

    Predicted band size: 193 kDa, 37 kDa, 45 kDa, 52 kDa, 54 kDa, 98 kDa

    Observed band size: 115 kDa, 193 kDa, 35 kDa, 37 kDa, 45 kDa, 52 kDa, 54 kDa

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com