AB187091

重组Anti-MLKL (phospho S358)抗体[EPR9514]

Name: MLKL(磷酸化S358)抗体[EPR9514] (ab187091)| Abcam中文官网
Brand: Abcam Limited
SKU: AB187091
Price: 1545 CNY
Availability: InStock
Rating: 4 (17 reviews)

Anti-MLKL (phospho S358) antibody [EPR9514]

RabMAb
Recombinant
20ul selling size
了解详情

(17 Reviews)

(283 Publications)

Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) is a rabbit monoclonal antibody detecting MLKL in Western Blot, IHC-P, Dot Blot. Suitable for Human.

- Biophysical QC for unrivalled batch-batch consistency
- Over 210 publications

查看别名

Mixed lineage kinase domain-like protein, hMLKL, MLKL

8 Images

Lab

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Blocking buffer and concentration : 5% NFDM/TBST, Diluting buffer and concentration : 5% NFDM /TBST, Exposure time : 1 minute

The lysate in this image is prepared by 1%SDS Hot Lysate buffer.

For Lysate preparation protocol, please refer to the protocol section of the website and/or here.

All lanes:

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1/1000 dilution

Lane 1:

Untreated HT-29 (human colorectal adenocarcinoma) whole cell lysates 20μg

Lane 2:

HT-29 (human colorectal adenocarcinoma) treated with TNF alpha+ Smac mimetic+ z-VAD whole cell lysates 20μg

Lane 3:

HT-29 (human colorectal adenocarcinoma) treated with TNF alpha+ Smac mimetic + z-VAD and phosphatase whole cell lysates 20μg.

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 54 kDa

Observed band size: 54 kDa

false

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

ab187091 at 1 : 250 staining MLKL (phospho S358) in Human skin tissue by immunohistochemistry (FFPE).

Antigen retrival required on FFPE tissue : HIER using 10mM Citrate buffer pH 6.0, see recommended HIER protocol

For additional IHC guideline, please see IHC resource page

IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

ab187091 at 1 : 250 staining MLKL (phospho S358) in Human melanoma tissue by immunohistochemistry (FFPE).

Antigen retrival required on FFPE tissue : HIER using 10mM Citrate buffer pH 6.0, see recommended HIER protocol

For additional IHC guideline, please see IHC resource page

AbReview64842****

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

All lanes:

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1000 Cells

Lane 1:

Untreated human hepatocyte cell lysate at 20 µg

Lane 2:

Treated (10 ng/ml TNF-a+100 nM Smac mimetic+20 µM z-VAD 6 h) human hepatocyte cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/5000 dilution

Predicted band size: 54 kDa

false

Data courtesy of an anonymous AbReview

Supplier Data

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Details on WB tested positive control samples : HT-29 cells were treated with the indicated stimuli for 8 hr and then harvested. The final concentrations of 20 ng/ml TNF-a, 100 nM Smac mimetic, and 20 μM z-VAD were used to induce necrosis.

The lysate in this image is prepared by 1%SDS Hot Lysate buffer.

For Lysate preparation protocol, please refer to the protocol section of the website and/or here.

All lanes:

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1/2000 dilution

Lane 1:

Untreated HT-29 lysate at 10 µg

Lane 2:

HT-29 cell lysate treated with TNF alpha+ Smac mimetic+ z-VAD at 10 µg

Secondary

All lanes:

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

Predicted band size: 54 kDa

false

Lab

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Sample : HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 μM z-VAD for 8 hours whole cell lysates 10 μg per lane.

Lane 1 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.12 μg/ml (Batch produced in 2016)
Lane 2 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.17 μg/ml (Batch produced in 2015)
Lane 3 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.12 μg/ml (GR212667 - batch produced in 2014)
Lane 4 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.16 μg/ml (The supernatant of the clone producing ab187091)
Lane 5 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.15 μg/ml (Batch produced in 2017)

SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Blocking and diluting buffer : 5% NFDM/TBST.

The lysate in this image is prepared by 1%SDS Hot Lysate buffer.

For Lysate preparation protocol, please refer to the protocol section of the website and/or here.

All lanes:

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091)

Predicted band size: 54 kDa

Observed band size: 54 kDa

false

Exposure time: 1min

Lab

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Blocking and diluting buffer : 5% NFDM/TBST.

For 1%SDS Hot Lysate preparation protocol, please refer to the protocol section of the website and/or here.

All lanes:

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1/5000 dilution

Lane 1:

HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hr. The lysate is directly prepared by 1xSDS loading buffer. at 20 µg

Lane 2:

HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 8 hr. The lysate is prepared by 1%SDS Hot Lysate buffer method. at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 54 kDa

Observed band size: 54 kDa

false

Exposure time: 3min

Lab

Dot Blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Dot blot analysis of MLKL peptides using ab187091 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated secondary antibody at 1/1000 dilution. Blocking and diluting buffer was 5% NFDM/TBST, exposure time 3 minuts.

Lane 1 : MLKL (pT357) phospho peptide

Lane 2 : MLKL (pS358) phospho peptide

Lane 3 : MLKL (pT357/pS358) phospho peptide

Lane 4 : MLKL non-phospho peptide

不同偶联物与剂型 (1)

Carrier free

Anti-MLKL (phospho S358) antibody [EPR9514] - BSA and Azide free

关键信息

宿主种属

Rabbit

克隆

Monoclonal

克隆号

EPR9514

亚型

IgG

不含载体蛋白

反应种属

Human

应用

IHC-P, Dot, WB

applications

免疫原

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

特异性

Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls.

FURTHER INFORMATION ON SPECIFICITY (Chinese Version) available under the product protocols section. This file includes key technical notes of experience when using this product.

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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/250 - 1/500", "IHCP-species-notes": "Not Suitable for Mouse and Rat Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000 - 1/2000", "WB-species-notes": "We recommend using 1% SDS Hot lysis method to prepare cell lysates.For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or <a data-tabindex-counter=\"1\" data-tabindex-value=\"none\" href=\"../../technical-resources/protocols/lysate-preparation-for-western-blot\" tabindex=\"-1\">here.</a>" }, "Mouse": { "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/250 - 1/500", "IHCP-species-notes": "Not Suitable for Mouse and Rat Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "1/1000 - 1/2000", "WB-species-notes": "We recommend using 1% SDS Hot lysis method to prepare cell lysates.For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or <a data-tabindex-counter=\"1\" data-tabindex-value=\"none\" href=\"../../technical-resources/protocols/lysate-preparation-for-western-blot\" tabindex=\"-1\">here.</a>" }, "Rat": { "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/250 - 1/500", "IHCP-species-notes": "Not Suitable for Mouse and Rat Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "1/1000 - 1/2000", "WB-species-notes": "We recommend using 1% SDS Hot lysis method to prepare cell lysates.For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or <a data-tabindex-counter=\"1\" data-tabindex-value=\"none\" href=\"../../technical-resources/protocols/lysate-preparation-for-western-blot\" tabindex=\"-1\">here.</a>" }, "Synthetic peptide - Human": { "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "" } } }

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产品详情

Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Dot Blot, IHC-P and WB.

Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) was first used in a scientific publication in 2012 and has been cited over 216 times in peer reviewed journals. It's performance in Western Blot in human samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) has 17 independent reviews from customers.

Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) specifically detects MLKL Phospho-S358 (UniProt ID: Q8NB16; Molecular weight: 55kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR9514 - ab208909.

Mixed lineage kinase domain-like protein (MLKL) is a key executor of necroptosis, a form of programmed cell death and its expression is associated with both tumor suppression and promotion, depending on the cancer type. Its role in necroptosis and other cellular processes makes it a potential prognostic marker and therapeutic target in oncology.

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

What are the advantages of a recombinant monoclonal antibody?
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.

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性能和储存信息

形式

Liquid

纯化工艺

Affinity purification Protein A

存储溶液

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

运输条件

Blue Ice

分装信息

Upon delivery aliquot

储存信息

Avoid freeze / thaw cycle

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补充信息

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

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.

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.

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产品实验方案

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

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靶点信息

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).

See full target information MLKL phospho S358

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文献 (283)

Recent publications for all applications. Explore the full list and refine your search

World journal of oncology 16:457-470 PubMed41030638

2025

Liquid Plasma Induces Necroptosis Without Inflammatory Responses in Head and Neck Cancer Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Jae Hoon Choi,Sungryeal Kim,Yun Sang Lee,Young Suk You,Jeon Yeob Jang,Yoo Seob Shin,Chul-Ho Kim

Communications biology 8:1384 PubMed41028907

2025

Casein kinase I isoforms contribute to platelet activation and thrombogenesis via RIPK3-MLKL signaling.

Applications

Unspecified application

Species

Unspecified reactive species

Vipin Singh,Mohammad Ekhlak,Susheel N Chaurasia,Debabrata Dash

Biology direct 20:98 PubMed40983928

2025

Necroptosis-driven T cell activation promotes IL-6-mediated PD-L1 upregulation in cholangiocarcinoma cells: IL-6 gene signature as a biomarker for chemo-immunotherapy response.

Applications

Unspecified application

Species

Unspecified reactive species

Thanpisit Lomphithak,Nattaya Duangthim,Sasiprapa Sonkaew,Siriporn Jitkaew

Cell research 35:840-858 PubMed40940518

2025

Soluble tissue factor generated by necroptosis-triggered shedding is responsible for thrombosis.

Applications

Unspecified application

Species

Unspecified reactive species

Peixing Wan,Swati Choksi,Yeon-Ji Park,Xin Chen,Jiong Yan,Sahar Foroutannejad,Zhaoshan Liu,Jichun Chen,Ross Lake,Chengyu Liu,Zheng-Gang Liu

Cell death discovery 11:413 PubMed40877228

2025

Discovery of Zharp1-163 as a dual inhibitor of ferroptosis and necroptosis for the treatment of inflammatory disorders and kidney injury.

Applications

Unspecified application

Species

Unspecified reactive species

Yuting Ji,Shujing Du,Jingjing Li,Haikuo Ma,Xinhui Wang,Yongjin Hao,Zhanhui Li,Haohao Lu,Hao Liu,Chengkui Yang,Xiaohu Zhang,Sudan He

Research (Washington, D.C.) 8:0823 PubMed40822127

2025

USP53 Drives Ethanol-Induced Myocardial Injury by Promoting K63 Deubiquitination-Dependent RIPK1 Activation at K377.

Applications

Unspecified application

Species

Unspecified reactive species

Jichen Pan,Xiaolin Liu,Xiao Li,Shanshan Wang,Yuliang Zhao,Chong Yuan,Dongdong Liu,Liyan Wang,Meng Zhang,Fengming Liu,Mei Zhang,Shen Dai

Frontiers in cell death 3: PubMed40809836

2025

ATF4-mediated expression of NOXA is critical for Necroptosis driven by Glucose Deprivation.

Applications

Unspecified application

Species

Unspecified reactive species

Sasiprapa Sonkaew,Ruwaida Rajna,Yeon-Ji Park,Jiong Yan,Zhaoshan Liu,Siriporn Jitkaew,Zheng-Gang Liu,Swati Choksi

Cell death discovery 11:345 PubMed40715038

2025

RIPK1 S213E mutant suppresses RIPK1-dependent cell death by preventing interactions with RIPK3 and CASP8.

Applications

Unspecified application

Species

Unspecified reactive species

Ning Nan,Hong Hu,Xinxin Zhu,Jia Liu,Feiyang Yuan,Zhijie Li,Huayi Wang

Frontiers in pharmacology 16:1586705 PubMed40458801

2025

Gambogic acid targets HSP90 to alleviate DSS-induced colitis via inhibiting the necroptosis of intestinal epithelial cells.

Applications

Unspecified application

Species

Unspecified reactive species

Yuanyuan Wang,Siqi Liu,Keyi Lu,Erping Xu,Zhibin Wang

Cell death & disease 16:426 PubMed40456737

2025

FDA-approved phensuximide inhibits RIPK1-dependent immunogenic cell death.

Applications

Unspecified application

Species

Unspecified reactive species

Byeong-Ju Kim,Sun Mi Hong,Hyun-Jin Noh,Jihye Kim,Su-Yeon Seon,Jeong-Eun Lee,Da-Hye Jeong,Ju-Mi Park,Sejeong Park,Sanghoon Lee,Jaewoo Kang,Dakeun Lee,Michael J Morgan,You-Sun Kim

View all publications

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websiteProtocolBooklet

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Abcam Product Promise

我们致力于为您的研究提供高质量的试剂，为您科研的每一步提供支持。若我们的产品未能达到预期性能，我们向您提供 Abcam Product Promise 保障。

详情请参阅我们的条款与条件。

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

重组Anti-MLKL (phospho S358)抗体[EPR9514]

Anti-MLKL (phospho S358) antibody [EPR9514]

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Western blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)

Dot Blot - Anti-MLKL (phospho S358) antibody [EPR9514] (AB187091)