|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
|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|
17563-1-AP targets RIP3 in WB, IP, IHC, IF, CoIP, ELISA applications and shows reactivity with human samples.
|Cited Reactivity||human, mouse, yeast, pig|
|Host / Isotype||Rabbit / IgG|
|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 ID (NCBI)||11035|
|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.|
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.
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