AB208430

HRP Anti-MLKL (phospho S345) antibody [EPR9515(2)]

RabMAb
Recombinant
What is this?

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(1 Publication)

Rabbit Recombinant Monoclonal MLKL phospho S345 antibody - conjugated to HRP. Suitable for Dot, WB and reacts with Synthetic peptide, Mouse samples. Cited in 1 publication.

View Alternative Names

Mixed lineage kinase domain-like protein, hMLKL, MLKL

2 Images

Lab

Western blot - HRP Anti-MLKL (phospho S345) antibody [EPR9515(2)] (AB208430)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - HRP Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab208430) at 1/4500 dilution

Lane 1:

Untreated L-929 (Mouse connective tissue fibroblast cell line) whole cell lysates at 15 µg

Lane 2:

L-929 (Mouse connective tissue fibroblast cell line) treated with 20 ng/ml TNF alpha,100nM Smac mimetic , and 20mM z-VAD for 8 hours whole cell lysates at 15 µg

Lane 3:

L-929 (Mouse connective tissue fibroblast cell line) treated with 20 ng/ml TNF alpha, 100nM Smac mimetic , and 20mM z-VAD for 8 hours whole cell lysates. Then the membrane was incubated with Alkaline phosphatase at 15 µg

Predicted band size: 54 kDa

Observed band size: 54 kDa

false

Exposure time: 1min

Lab

Dot Blot - HRP Anti-MLKL (phospho S345) antibody [EPR9515(2)] (AB208430)

Dot Blot analysis of

Lane 1 : MLKL (phospho S345) phospho peptide and

Lane 2 : MLKL Non-phospho peptide labeling MLKL (phospho S345)

with ab208430 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. Exposure time : 3 minutes.

Unconjugated

Anti-MLKL (phospho S345) antibody [EPR9515(2)]
Carrier free

Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR9515(2)

Isotype

IgG

Conjugation

HRP

Excitation/Emission

Carrier free

Reacts with

Mouse

Applications

Dot, WB

applications

Immunogen

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

Specificity

MLKL pS345 is a trigger for necroptosis. It is only detectable in infection/cellular damaged (PMID:29229989) or aging tissue (PMID: 28807105) but not in normal tissues.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "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": { "Mouse": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/4500", "WB-species-notes": "<p></p>" }, "Synthetic peptide": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "" } } }

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Product details

This antibody was developed through collaboration with the lab of Xiaodong Wang at the National Institute of Biological Sciences, Beijing.

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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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.4 Preservative: 0.1% Proclin 300 Solution Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA

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|Store in the dark

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Supplementary information

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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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

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 S345

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Publications (1)

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

Frontiers in bioengineering and biotechnology 10:885105 PubMed35646869

2022

Regulating Acidosis and Relieving Hypoxia by Platelet Membrane-Coated Nanoparticle for Enhancing Tumor Chemotherapy.

Applications

Unspecified application

Species

Unspecified reactive species

Xingyu Luo,Jian Cao,Jianming Yu,Dongqing Dai,Wei Jiang,Yahui Feng,Yong Hu

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

HRP Anti-MLKL (phospho S345) antibody [EPR9515(2)]

Western blot - HRP Anti-MLKL (phospho S345) antibody [EPR9515(2)] (AB208430)

Dot Blot - HRP Anti-MLKL (phospho S345) antibody [EPR9515(2)] (AB208430)

Related conjugates and formulations (2)

Key facts

Host species

Clonality

Clone number

Isotype

Conjugation

Excitation/Emission

Carrier free

Reacts with

Applications

Immunogen

Specificity

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

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