Rabbit Recombinant Monoclonal RIP3 phospho S232 antibody. Carrier free. Suitable for ELISA, Dot, WB and reacts with Synthetic peptide, Mouse samples.
Images
![Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (AB240336), expandable thumbnail](https://content.abcam.com/products/images/rip3-phospho-s232-antibody-epr9516n-25-bsa-and-azide-free-ab240336--dot-blot-img13255.jpg/_jcr_content/renditions/webp-475.webp "Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (AB240336)")
![ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (AB240336), expandable thumbnail](https://content.abcam.com/products/images/rip3-phospho-s232-antibody-epr9516n-25-bsa-and-azide-free-ab240336--elisa-img82633.jpg/_jcr_content/renditions/webp-475.webp "ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (AB240336)")
![Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (AB240336), expandable thumbnail](https://content.abcam.com/products/images/rip3-phospho-s232-antibody-epr9516n-25-bsa-and-azide-free-ab240336--western-blot-img433179.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (AB240336)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| ELISA | Dot | WB | |
|---|---|---|---|
| Mouse | Expected | Expected | Tested |
| Synthetic peptide | Tested | Tested | Not recommended |
ELISA
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Dot
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
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
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 S232 antibody. Carrier free. Suitable for ELISA, Dot, WB and reacts with Synthetic peptide, Mouse samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR9516(N)-25
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- Do Not Freeze
Notes
ab240336 is the carrier-free version of Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] ab195117.
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.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our 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.
3 product images
![Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336), expandable thumbnail](https://content.abcam.com/products/images/rip3-phospho-s232-antibody-epr9516n-25-bsa-and-azide-free-ab240336--dot-blot-img13255.jpg "Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336)")
Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336)
Primary antibody: Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] ab195117 at 1:1000 dilution (1.6μg/ml), secondary antibody: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:1000 dilution, Blocking and dilution buffer: 5% NFDM/TBST, Lane 1: RIP3 (phospho S232) phospho peptide, Lane 2: RIP3 Non-phospho peptide, Exposure time: 3 minutes.
We observed weak binding to the non-phospho peptide at very high concentrations.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] ab195117).
![ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336), expandable thumbnail](https://content.abcam.com/products/images/rip3-phospho-s232-antibody-epr9516n-25-bsa-and-azide-free-ab240336--elisa-img82633.jpg "ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336)")
ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336)
Serially diluted Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] ab195117 was bound to immobilized phospho or non-phospho (control) peptide demonstrating minimal cross-reactivity to the non-phosphorylated residue. Antigen - RIP3 (phospho S232) phospho peptide;RIP3 non-phospho peptide at 250 ng/ml. Primary antibody concentration range: 0 - 500 ng/ml. Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1:2500.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] ab195117).
![Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336), expandable thumbnail](https://content.abcam.com/products/images/rip3-phospho-s232-antibody-epr9516n-25-bsa-and-azide-free-ab240336--western-blot-img433179.jpg "Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336)")
Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] ab195117).
Blocking and diluting buffer: 5% NFDM/TBST.
Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 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 S232) antibody [EPR9516(N)-25] (Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] ab195117) 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
SecondaryAll 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
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
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