Description
Product Usage Information
Western Blotting: 1:1000
Immunofluorescence (Immunocytochemistry): 1:200 - 1:400
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 50433 .
Protocol
Available protocols: Western Blotting, Immunofluorescence (Immunocytochemistry)
Specificity / Sensitivity
Phospho-RIP3 (Thr231/Ser232) (E7S1R) Rabbit Monoclonal Antibody recognizes endogenous levels of RIP3 protein only when phosphorylated at Thr231/Ser232. This antibody may not recognize RIP3 when only singly phosphorylated at Thr231 or Ser232.
Species Reactivity: Mouse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Thr231/Ser232 of mouse RIP3 protein.
Background
The receptor-interacting protein (RIP) family of serine-threonine kinases (RIP, RIP2, RIP3, and RIP4) are important regulators of cellular stress that trigger pro-survival and inflammatory responses through the activation of NF-κB, as well as pro-apoptotic pathways (1). In addition to the kinase domain, RIP contains a death domain responsible for interaction with the death domain receptor Fas and recruitment to TNF-R1 through interaction with TRADD (2,3). RIP-deficient cells show a failure in TNF-mediated NF-κB activation, making the cells more sensitive to apoptosis (4,5). RIP also interacts with TNF-receptor-associated factors (TRAFs) and can recruit IKKs to the TNF-R1 signaling complex via interaction with NEMO, leading to IκB phosphorylation and degradation (6,7). Overexpression of RIP induces both NF-κB activation and apoptosis (2,3). Caspase-8-dependent cleavage of the RIP death domain can trigger the apoptotic activity of RIP (8). Receptor-interacting protein 3 (RIP3) was originally found to interact with RIP and the TNF receptor complex to induce apoptosis and activation of NF-κB (9,10). It has subsequently been shown that the association between RIP and RIP3 is a key component of a signaling pathway that results in programmed necrosis (necroptosis), a necrotic-like cell death induced by TNF in the presence of caspase inhibitors (11-13). RIP3 is phosphorylated at Ser227 and targets the phosphorylation of mixed lineage kinase domain-like protein (MLKL), which is critical for necroptosis (14). In mice, RIP3 is phosphorylated at Thr231 and Ser232, leading to association with MLKL and necroptosis (15).
Alternate Names
2610528K09Rik; AW107945; mRIP3; receptor interacting protein 3; Receptor-interacting protein 3; receptor-interacting serine-threonine kinase 3; Receptor-interacting serine/threonine-protein kinase 3; RIP-3; RIP-like protein kinase 3; Rip3; Ripk3
