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- KD/KO Validated
RIP3 Polyclonal antibody
RIP3 Polyclonal Antibody for IHC, IP, WB, ELISA
Host / Isotype
Rabbit / IgG
Reactivity
human and More (3)
Applications
WB, IP, IHC, IF, CoIP, ELISA
Conjugate
Unconjugated
Cat no : 17563-1-AP
Synonyms
Validation Data Gallery
Tested Applications
| Positive WB detected in | HT-29 cells, HeLa cells, SW 1990 cells, Jurkat cells, THP-1 cells, SGC-7901 cells |
| Positive IP detected in | SW 1990 cells |
| Positive IHC detected in | human pancreas tissue Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0 |
Recommended dilution
| Application | Dilution |
|---|---|
| Western Blot (WB) | WB : 1:1000-1:4000 |
| Immunoprecipitation (IP) | IP : 0.5-4.0 ug for IP and 1:200-1:1000 for WB |
| Immunohistochemistry (IHC) | IHC : 1:100-1:400 |
| Sample-dependent, check data in validation data gallery | |
Published Applications
| KD/KO | See 1 publications below |
| WB | See 89 publications below |
| IHC | See 10 publications below |
| IF | See 12 publications below |
| IP | See 6 publications below |
| CoIP | See 2 publications below |
Product Information
17563-1-AP targets RIP3 in WB, IP, IHC, IF, CoIP, ELISA applications and shows reactivity with human samples.
| Tested Reactivity | human |
| Cited Reactivity | human, mouse, yeast, pig |
| Host / Isotype | Rabbit / IgG |
| Class | Polyclonal |
| Type | Antibody |
| Immunogen | RIP3 fusion protein Ag11759 |
| Full Name | receptor-interacting serine-threonine kinase 3 |
| Calculated molecular weight | 518 aa, 57 kDa |
| Observed molecular weight | 57 kDa |
| GenBank accession number | BC062584 |
| Gene symbol | RIPK3 |
| Gene ID (NCBI) | 11035 |
| Conjugate | Unconjugated |
| Form | Liquid |
| Purification Method | Antigen affinity purification |
| Storage Buffer | PBS with 0.02% sodium azide and 50% glycerol pH 7.3. |
| Storage Conditions | Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA. |
Background Information
Receptor-interacting protein 3 (RIP3, also known as RIPK3) is a serine-threonine protein involved in the regulation of inflammatory signaling and cell death. RIPK3, also named as RIP3, a Ser/Thr kinase of RIP (Receptor Interacting Protein) family, is a nucleocytoplasmic shuttling protein and its unconventional nuclear localization signal (NLS, 442-472 aa) is sufficient to trigger apoptosis in the nucleus (PMID: 18533105). It has 3 isoforms produced by alternative splicing.
What is the molecular weight of RIP3? Is RIP3 post-translationally modified?
The molecular weight of RIP3 is 57 kDa. During the induction of necroptosis, RIP3 migrates slower in SDS-PAGE due to its phosphorylation (PMID: 19524512). Additionally, RIP3 can be a subject of poly-ubiquitination, when targeted for degradation.
Are there any splice isoforms of RIP3?
Apart from full-length RIP3, there are two reported splice isoforms of RIP3: RIP3β and RIP3γ, and 28 and 25 kDa, respectively (PMID: 15896315).
What is the subcellular localization of RIP3?
RIP3 can shuttle between the cytoplasm and the nucleus. Although RIP3 forms necrosomes in the cytoplasm, a recent study suggests that the phosphorylation of RIP3, required for necroptosis, may also occur in the nucleus (PMID: 30271893). Additionally, the induction of necrosis by reactive oxygen species can cause transient translocation of RIP3 to mitochondria (PMID: 25206339).
What is the role of RIP3 in cell death (necroptosis)?
Activation of RIP3 kinase is required for the induction of necroptosis (PMID: 19524512, 19524513 and 19498109). Activation of RIP3 can be induced by interferons, death ligands, or by Toll-like receptors in response to pathogens. That leads to the phosphorylation of RIP3 and the formation of a β-amyloid-like protein complex. Phosphorylated RIP3 acts downstream by phosphorylation of MLKL (PMID: 30131615).
How to study necroptosis in a cell-based system?
The choice of the cell line is important. Many commonly used immortalized cell types are derived from cancers and may have very low RIL3 expression level (PMID: 25952668). Those cell lines are not going to be responsive to necrotic stimuli. A few of the examined cell types have high RIP3 levels: Jurkat, CCRF-CEM, U937, L929 cells, and mouse embryonic fibroblasts (PMID: 19524512). Good necroptosis readouts reflect an increased level of RIP3 protein and its phosphorylation.
Protocols
| Product Specific Protocols | |
|---|---|
| WB protocol for RIP3 antibody 17563-1-AP | Download protocol |
| IHC protocol for RIP3 antibody 17563-1-AP | Download protocol |
| IP protocol for RIP3 antibody 17563-1-AP | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
| Species | Application | Title |
|---|---|---|
Acta Pharm Sin B Ligand-based substituent-anchoring design of selective receptor-interacting protein kinase 1 necroptosis inhibitors for ulcerative colitis therapy | ||
ACS Appl Mater Interfaces Enhancing Tumor Therapy of Fe(III)-Shikonin Supramolecular Nanomedicine via Triple Ferroptosis Amplification | ||
Front Immunol N-acetyl-L-cysteine alleviated the oxidative stress-induced inflammation and necroptosis caused by excessive NiCl2 in primary spleen lymphocytes | ||
Environ Pollut Biotransformation of graphene oxide within lung fluids could intensify its synergistic biotoxicity effect with cadmium by inhibiting cellular efflux of cadmium. | ||
Life Sci TNF-α increases Staphylococcus aureus-induced death of human alveolar epithelial cell line A549 associated with RIP3-mediated necroptosis. | ||
Cell Mol Life Sci Thioredoxin reductase 3 suppression promotes colitis and carcinogenesis via activating pyroptosis and necrosis. |
