Proteintech, 17563-1-AP, RIPK3 Polyclonal antibody

Size: 20ul / 150ul The RIPK3 (17563-1-AP) by Proteintech is a Polyclonal antibody targeting RIPK3 in WB, IHC, IF/ICC, IP, ELISA applications with reactivity to human samples 17563-1-AP targets RIPK3 in WB, IHC, IF/ICC, IP, CoIP, ELISA applications and shows reactivity with human samples. Tested Applications Positive WB detected in: HT-29 cells, Jurkat cells, SW 1990 cells, THP-1 cells, SGC-7901 cells Positive IP detected in: SW 1990 cells Positive IHC detected in: human pancreas tissue, human placenta tissueNote: suggested antigen retrieval withTE buffer pH 9.0;(*) Alternatively, antigen retrieval may be performed withcitrate buffer pH 6.0 Positive IF/ICC detected in: HT-29 cells Recommended dilution Western Blot (WB): WB : 1:1000-1:4000 Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate Immunohistochemistry (IHC): IHC : 1:100-1:400 Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500 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. Specification Tested Reactivity: human Cited Reactivity: human, yeast Host / Isotype: Rabbit / IgG Class: Polyclonal Type: Antibody Immunogen: CatNo: Ag11759 Product name: Recombinant human RIPK3 protein Source: e coli. -derived, PGEX-4T Tag: GST Domain: 1-289 aa of BC062584 Sequence: MSCVKLWPSGAPAPLVSIEELENQELVGKGGFGTVFRAQHRKWGYDVAVKIVNSKAISREVKAMASLDNEFVLRLEGVIEKVNWDQDPKPALVTKFMENGSLSGLLQSQCPRPWPLLCRLLKEVVLGMFYLHDQNPVLLHRDLKPSNVLLDPELHVKLADFGLSTFQGGSQSGTGSGEPGGTLGYLAPELFVNVNRKASTASDVYSFGILMWAVLAGREVELPTEPSLVYEAVCNRQNRPSLAELPQAGPETPGLEGLKELMQLCWSSEPKDRPSFQECLPKTDEVFQM Predict reactive species Full Name: receptor-interacting serine-threonine kinase 3 Calculated Molecular Weight: 518 aa, 57 kDa Observed Molecular Weight: 57-70 kDa GenBank Accession Number: BC062584 Gene Symbol: RIPK3 Gene ID (NCBI): 11035 RRID: AB_2178659 Conjugate: Unconjugated Form: Liquid Purification Method: Antigen affinity purification UNIPROT ID: Q9Y572 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 -20 o C storage. 20ul sizes contain 0.1% BSA.