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AB255705

Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20]

  • 20ul selling size
  • RabMAb
  • Recombinant
  • KO Validated
  • What is this?

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(3 Publications)

Rabbit Recombinant Monoclonal RIP3 phospho S164 + T165 antibody. Suitable for Dot, WB, IHC-Fr and reacts with Synthetic peptide - Mouse, Mouse, Recombinant fragment - Mouse samples. Cited in 3 publications.

View Alternative Names

Rip3, Ripk3, Receptor-interacting serine/threonine-protein kinase 3, RIP-like protein kinase 3, Receptor-interacting protein 3, RIP-3, mRIP3

4 Images
Immunohistochemistry (Frozen sections) - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)
  • IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse ovary tissue labeling RIP3 (phospho S164 + T165) with ab255705 at 1/50 dilution (10 μg/mL) followed by an Alexa Fluor® 555 Donkey anti-Rabbit secondary at 1/100 dilution (Green). Image was kindly provided by collaborator. Positive staining on wild-type mouse ovary, while no staining on RIP3 KO mouse ovary is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is an Alexa Fluor® 555 Donkey anti-Rabbit secondary at 1/100 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)
  • WB

Lab

Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Image was kindly provided by collaborator.

Exposure time : 60 seconds.

All lanes:

Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (ab255705) at 1/500 dilution

Lane 1:

Mouse ovary(4 months) tissue lysate at 20 µg

Lane 2:

Mouse ovary(8 months) tissue lysate at 20 µg

Lane 3:

Mouse ovary(12 months) tissue lysate at 20 µg

Lane 4:

Mouse ovary(16 months) tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/1000 dilution

Predicted band size: 57 kDa

false

Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)
  • WB

Lab

Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lysates were kindly provided by collaborator.

Exposure time : 3 seconds.

All lanes:

Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (ab255705) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) transfected with RIP3 mutant (S164A/T165A) expression vector, treated with DOX and Z-VAD whole cell lysate at 20 µg

Lane 2:

MCF7 transfected with RIP3 (WT) expression vector, treated with DOX and Z-VAD whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 57 kDa

false

Dot Blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)
  • Dot

Supplier Data

Dot Blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705)

Dot blot analysis of RIP3 (phospho S164 + T165) using ab255705 at 1/1000 (0.46 μg/mL) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

Lane 1 : RIP3 (phospho S164 + T165) peptide
Lane 2 : RIP3 (phospho S164 + T165) peptide
Lane 3 : RIP3 non-phospho peptide
Lane 4 : RIP3 (phospho S164) peptide
Lane 5 : RIP3 (phospho T165) peptide

Exposure time : 3 minutes

Blocking and diluting buffer and concentration : 5% NFDM/TBST

  • Carrier free

    Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR23660-20

Isotype

IgG

Carrier free

No

Reacts with

Mouse

Applications

WB, IHC-Fr, Dot

applications

Immunogen

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

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Primary Antibodies

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Primary Antibodies

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Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
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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"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IHCFr" : {"fullname" : "Immunohistochemistry (Frozen sections)", "shortname":"IHC-Fr"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Mouse": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/500", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IHCFr-species-checked": "testedAndGuaranteed", "IHCFr-species-dilution-info": "1/50", "IHCFr-species-notes": "<p></p>" }, "Rat": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Recombinant fragment - Mouse": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/500", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Synthetic peptide - Mouse": { "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": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" } } }
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Product details

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

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

'Receptor-interacting protein kinase 3' (RIP3) also known as RIPK3 is a serine/threonine-protein kinase with a molecular weight of approximately 57 kDa. Mechanically it contains a kinase domain that allows it to phosphorylate specific substrates which is important for mediating its function within the cell. RIP3 is expressed in various tissues with notable presence in the spleen heart and adipose tissue. The protein localizes predominantly in the cytoplasm where it interacts with other cellular proteins to initiate downstream signaling events.
Biological function summary

RIP3 facilitates the execution of necroptosis a form of programmed cell death distinct from apoptosis. It becomes activated upon binding with RIP1 forming a necrosome complex that is essential for this pathway. This complex promotes phosphorylation events that subsequently lead to membrane rupture and cell death. Apart from its role in necroptosis RIP3 also engages in metabolic regulation processes linking energy status and cell death under conditions of stress.

Pathways

RIP3 is a principal component of the necroptotic pathway interacting closely with RIP1 to trigger cell death in conditions where caspase activation is inhibited. Alternatively it integrates into metabolic pathways participating in sensing and responding to changes in cellular energy states. The interplay between RIP3 and RIP1 within these pathways illustrates their shared involvement in maintaining cellular homeostasis and triggering cell death when necessary.

RIP3 has significant implications for conditions involving excessive or dysfunctional cell death such as inflammatory diseases and reperfusion injury. The necroptotic activity of RIP3 can exacerbate inflammation by promoting the release of pro-inflammatory factors upon cell death. Furthermore during ischemia-reperfusion injury increased RIP3 activity in conjunction with MLKL another necroptosis-associated protein contributes to tissue damage highlighting its potential as a therapeutic target for reducing cell death-related tissue damage.
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Product protocols

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

Serine/threonine-protein kinase that activates necroptosis and apoptosis, two parallel forms of cell death (PubMed : 27321907, PubMed : 27746097, PubMed : 27917412, PubMed : 28607035, PubMed : 32200799, PubMed : 32296175). Necroptosis, a programmed cell death process in response to death-inducing TNF family members, is triggered by RIPK3 following activation by ZBP1 (PubMed : 19590578, PubMed : 22423968, PubMed : 24012422, PubMed : 24019532, PubMed : 24095729, PubMed : 24557836, PubMed : 27321907, PubMed : 27746097, PubMed : 27819681, PubMed : 27819682, PubMed : 32200799, PubMed : 32296175). Activated RIPK3 forms a necrosis-inducing complex and mediates phosphorylation of MLKL, promoting MLKL localization to the plasma membrane and execution of programmed necrosis characterized by calcium influx and plasma membrane damage (PubMed : 24813849, PubMed : 24813850, PubMed : 27321907). In addition to TNF-induced necroptosis, necroptosis can also take place in the nucleus in response to orthomyxoviruses infection : 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 (PubMed : 32200799, PubMed : 32296175). Also regulates apoptosis : apoptosis depends on RIPK1, FADD and CASP8, and is independent of MLKL and RIPK3 kinase activity (PubMed : 27321907). Phosphorylates RIPK1 : RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation (By similarity). In some cell types, also able to restrict viral replication by promoting cell death-independent responses (PubMed : 30635240). In response to flavivirus infection in neurons, promotes a cell death-independent pathway that restricts viral replication : together with ZBP1, promotes a death-independent transcriptional program that modifies the cellular metabolism via up-regulation expression of the enzyme ACOD1/IRG1 and production of the metabolite itaconate (PubMed : 30635240). Itaconate inhibits the activity of succinate dehydrogenase, generating a metabolic state in neurons that suppresses replication of viral genomes (PubMed : 30635240). RIPK3 binds to and enhances the activity of three metabolic enzymes : GLUL, GLUD1, and PYGL (By similarity). These metabolic enzymes may eventually stimulate the tricarboxylic acid cycle and oxidative phosphorylation, which could result in enhanced ROS production (By similarity).
See full target information Ripk3 phospho S164 + T165
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Publications (3)

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

Drug design, development and therapy 19:1909-1926 PubMed40098903

2025

Design of a Highly Active Peptide Inhibitor of Farnesyltransferase and Its Protective Effect Against Acute Liver Failure.

Applications

Unspecified application

Species

Unspecified reactive species

Chun-Lian Huang,Hang-Shuai Qu,A-Li Li,Chen-Qian Ying,Hui Shao,Yong-Zhi Tang,Hua-Zhong Chen,Tao-Hsin Tung,Jian-Sheng Zhu

Frontiers in pharmacology 15:1436013 PubMed39329120

2024

Inhibition of the RIP3/MLKL/TRPM7 necroptotic pathway ameliorates diabetes mellitus-induced erectile dysfunction by reducing cell death, fibrosis, and inflammation.

Applications

Unspecified application

Species

Unspecified reactive species

Lipan Niu,Pei Yang,Bingbing Zhu,Xiufang Jin,Chengxia Yang,Xijia Zhang,Yulian Liu,Rui Zhang,Fengxia Liu

eLife 10: PubMed34029184

2021

A phosphorylation of RIPK3 kinase initiates an intracellular apoptotic pathway that promotes prostaglandin-induced corpus luteum regression.

Applications

IHC-Fr

Species

Mouse

Dianrong Li,Jie Chen,Jia Guo,Lin Li,Gaihong Cai,She Chen,Jia Huang,Hui Yang,Yinhua Zhuang,Fengchao Wang,Xiaodong Wang
View all publications
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Product promise

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For full details, please see our Terms & Conditions

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