Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
- Datasheet
- References (8)
- Protocols
Overview
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Product nameAnti-RIP3 (phospho S232) antibody [EPR9516(N)-25]
See all RIP3 primary antibodies -
DescriptionRabbit monoclonal [EPR9516(N)-25] to RIP3 (phospho S232)
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Host speciesRabbit
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Tested applicationsSuitable for: ELISA, WB, Dot blotmore details
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Species reactivityReacts with: Mouse
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Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Mouse RIP3 aa 200-300 (phospho S232). The exact sequence is proprietary.
Database link: Q9QZL0 -
Positive control
- WB: L-929 whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 8 h and then harvested; Untreated L-929 whole cell lysate; TSE (WT) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested; TSE (KO MLKL) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested.
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General 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 rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEPR9516(N)-25
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
Our Abpromise guarantee covers the use of ab195117 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ELISA | Use at an assay dependent concentration. | |
| WB | 1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa). | |
| Dot blot | Use at an assay dependent concentration. |
Target
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FunctionPromotes apoptosis.
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Tissue specificityDetected at lower levels in heart, placenta, lung and kidney. Isoform 3 is significantly increased in colon and lung cancers.
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Sequence similaritiesBelongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
Contains 1 protein kinase domain. -
Post-translational
modificationsAutophosphorylated. -
Cellular localizationCytoplasm.
- Information by UniProt
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Database links
- Entrez Gene: 56532 Mouse
- SwissProt: Q9QZL0 Mouse
- Unigene: 46612 Mouse
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Alternative names
- Receptor interacting protein 3 antibody
- Receptor interacting serine threonine kinase 3 antibody
- Receptor interacting serine/threonine protein kinase 3 antibody
see all
Images
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![Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)](https://www.abcam.com/ps/products/195/ab195117/Images/ab195117-284907-WB2v2.jpg)
Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)All lanes : Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117) at 1/10000 dilution
Lane 1 : Untreated L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
Lane 2 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate treated with phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)](https://www.abcam.com/ps/products/195/ab195117/Images/ab195117-257717-195117WBc.jpg)
Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)All lanes : Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117) at 1/2000 dilution
Lane 1 : Untreated TSE (WT) whole cell lysate
Lane 2 : TSE (WT) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested
Lane 3 : Untreated TSE (KO RIP3) whole cell lysate
Lane 4 : TSE (KO RIP3) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested
Lane 5 : Untreated TSE (KO MLKL) whole cell lysate
Lane 6 : TSE (KO MLKL) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Lysates were provided by Xiaodong Wang, National Institute of Biological Sciences, Beijing
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![Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)](https://www.abcam.com/ps/products/195/ab195117/Images/ab195117-258254-image-1.jpg)
Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)Primary antibody: ab195117 at 1:1000 dilution (1.6μg/ml), secondary antibody: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (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.
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![ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)](https://www.abcam.com/ps/products/195/ab195117/Images/ab195117-258156-image-1.jpg)
ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)Serially diluted 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.
Protocols
References
This product has been referenced in:
- Zechendorf E et al. Heparan Sulfate Induces Necroptosis in Murine Cardiomyocytes: A Medical-In silico Approach Combining In vitro Experiments and Machine Learning. Front Immunol 9:393 (2018). WB ; Mouse . Read more (PubMed: 29616016) »
- Fuchslocher Chico J et al. The enhanced susceptibility of ADAM-17 hypomorphic mice to DSS-induced colitis is not ameliorated by loss of RIPK3, revealing an unexpected function of ADAM-17 in necroptosis. Oncotarget 9:12941-12958 (2018). Read more (PubMed: 29560122) »
