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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 1 publication.

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Images

Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705), expandable thumbnail
  • Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705), expandable thumbnail
  • Immunohistochemistry (Frozen sections) - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705), expandable thumbnail
  • Dot Blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (AB255705), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
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
DotWBIHC-PIHC-Fr
Human
Not recommended
Not recommended
Not recommended
Not recommended
Mouse
Expected
Tested
Not recommended
Tested
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Recombinant fragment - Mouse
Not recommended
Tested
Not recommended
Not recommended
Synthetic peptide - Mouse
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Synthetic peptide - Mouse
Dilution info
1/1000
Notes

-

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Recombinant fragment - Mouse, Human, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Recombinant fragment - Mouse
Dilution info
1/500
Notes

-

Species
Mouse
Dilution info
1/500
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Mouse, Human, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human, Rat, Synthetic peptide - Mouse, Recombinant fragment - Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/50
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Mouse, Recombinant fragment - Mouse, Human, Rat
Dilution info
-
Notes

-

Target data

Function

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-alpha 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).

Alternative names

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

Recommended products

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 1 publication.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR23660-20
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

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

'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.

Associated diseases and disorders

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.

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.

4 product images

  • Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (ab255705), expandable thumbnail

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

    Predicted band size: 57 kDa

  • Western blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (ab255705), expandable thumbnail

    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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

    Predicted band size: 57 kDa

  • Immunohistochemistry (Frozen sections) - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (ab255705), expandable thumbnail

    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).

  • Dot Blot - Anti-RIP3 (phospho S164 + T165) antibody [EPR23660-20] (ab255705), expandable thumbnail

    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 (Goat Anti-Rabbit IgG H&L (HRP) 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

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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'.

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