CST, 83613S, Phospho-RIP (Ser321) Antibody

Polyclonal Antibody for studying RIPK1 (Ser321) phosphate mouse. Validated for Western Blotting. Highly specific and rigorously validated in-house, Phospho-RIP (Ser321) Antibody (CST #83613) is ready to ship. Product Usage Information Western Blotting: 1:1000 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting Specificity / Sensitivity Phospho-RIP (Ser321) Antibody recognizes endogenous levels of RIP protein only when phosphorylated at Ser321. Species Reactivity: Mouse Source / Purification Polyclonal antibodies are produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser321 of mouse RIP protein. Antibodies are purified by protein A and peptide affinity chromatography. 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). Necroptosis, a regulated pathway for necrotic cell death, is triggered by a number of inflammatory signals, including cytokines in the tumor necrosis factor (TNF) family, pathogen sensors such as toll-like receptors (TLRs), and ischemic injury (9,10). The process is negatively regulated by caspases and is initiated through a complex containing the RIP and RIP3 kinases, typically referred to as the necrosome. Necroptosis is inhibited by a small molecule inhibitor of RIP, necrostatin-1 (Nec-1) (11). Research studies show that necroptosis contributes to a number of pathological conditions, and Nec-1 has been shown to provide neuroprotection in models such as ischemic brain injury (12). RIP is phosphorylated at several sites within the kinase domain that are sensitive to Nec-1, including Ser14, Ser15, Ser161, and Ser166 (13). RIP is also phosphorylated at Ser321(mouse)/Ser320(human) by MAPKAPK-2 (MK-2) and TAK1 in response to inflammatory signals such as TNF-α and LPS (14-17). Phosporylation at this site suppresses RIP mediated apoptosis by inhibiting its interaction with FADD and caspase-8 (14-17). Alternate Names Cell death protein RIP; D330015H01Rik; receptor (TNFRSF)-interacting serine-threonine kinase 1; Receptor-interacting protein 1; Receptor-interacting serine/threonine-protein kinase 1; Rinp; Rip; RIP-1; Rip1; Ripk1; Serine/threonine-protein kinase RIP Specification REACTIVITY: M SENSITIVITY: Endogenous MW (kDa): 78 SOURCE: Rabbit