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Rabbit Recombinant Monoclonal RIP3 phospho S232 + T231 antibody. Suitable for Dot, WB and reacts with Synthetic peptide, Mouse samples. Cited in 21 publications.

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Images

Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (AB222320), expandable thumbnail
  • Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (AB222320), expandable thumbnail
  • Dot Blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (AB222320), expandable thumbnail
  • Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (AB222320), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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
DotWB
Mouse
Expected
Tested
Synthetic peptide
Tested
Not recommended

Tested
Tested

Species

Synthetic peptide

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide

Dilution info

-

Notes

-

Associated Products

Select an associated product type

8 products for Alternative Product

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

Recommended products

Rabbit Recombinant Monoclonal RIP3 phospho S232 + T231 antibody. Suitable for Dot, WB and reacts with Synthetic peptide, Mouse samples. Cited in 21 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR19403-52

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:
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4 product images

  • Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320), expandable thumbnail

    Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Cells were lysed in 4% SDS buffer (4% SDS,10% 2-ME, 20% glycerol, 0.004% Bromophenol blue, Tris-HCl).

    The images were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.

    All lanes: Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320) at 1/500 dilution

    Lane 1: Untreated L-929 (mouse connective tissue fibroblast cell line) whole cell lysate at 20 µg

    Lane 2: L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 10 ng/ml TNF-a and 20 µM z-VAD for 2 hours at 20 µg

    Lane 3: L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 10 ng/ml TNF-a and 20 µM z-VAD for 3.5 hours at 20 µg

    Lane 4: L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours at 20 µg

    Lane 5: L-929 (mouse connective tissue fibroblast cell line) whole cell lysate with RIP3 KO cells treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/5000 dilution

    Predicted band size: 57 kDa

    Exposure time: 60s

  • Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320), expandable thumbnail

    Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The higher MW bands in Lanes 1 and 2 are extra bands.

    The lysates were prepared following the 1% SDS Hot Lysate buffer method.

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

    All lanes: Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320) at 1/1000 dilution

    Lane 1: Untreated L-929 (mouse connective tissue fibroblast cell line) whole cell lysate at 10 µg

    Lane 2: L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours at 10 µg

    Lane 3: L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours, then treated with alkaline phosphatase for 1 hour at 10 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

    Observed band size: 53 kDa

    Exposure time: 30s

  • Dot Blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320), expandable thumbnail

    Dot Blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320)

    Dot blot analysis of RIP3 (phospho T231 + S232) labeled with ab222320 at 1/1000 dilution.

    Lane 1: RIP3 (phospho T231 + S232) peptide (aa229-239);

    Lane 2: RIP3 (phospho T231 + S232) peptide (aa223-236);

    Lane 3: RIP3 (phospho T231) peptide (aa223-239);

    Lane 4: RIP3 (phospho S232) peptide (aa223-239);

    Lane 5: RIP3 non-phospho peptide (aa223-239).

    Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320), expandable thumbnail

    Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320)

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

    Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 GAPDH was used as a loading control at 1/1000000 dilution.

    We recommend involving downstream protein p-MLKL as a control to validate the stimulation of p-RIP3.

    Lanes 1 - 4: Western blot - Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320) at 1/1000 dilution

    Lanes 5 - 8: Western blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (Anti-MLKL (phospho S345) antibody [EPR9515(2)] ab196436) at 1/1000 dilution

    Lanes 1, 3, 5 and 7: Untreated L-929 (mouse connective tissue fibroblast cell line) whole cell lysate at 20 µg

    Lanes 2 and 6: L-929 (mouse connective tissue fibroblast cell line) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 μM z-VAD for 8 hours whole cell lysate at 20 µg

    Lanes 4 and 8: L-929 (mouse connective tissue fibroblast cell line) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 μM z-VAD for 6 hours 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, 54 kDa

    Observed band size: 53 kDa

    Exposure time: 180s

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Product protocols

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

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