Description
Product Usage Information
Western Blotting: 1:1000
Immunofluorescence (Immunocytochemistry): 1:400
Flow Cytometry (Fixed/Permeabilized): 1:200 - 1:800
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 # 96323 .
Protocol
Available protocols: Western Blotting, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized)
Specificity / Sensitivity
Phospho-RIP (Ser166) (D8I3A) Rabbit Monoclonal Antibody recognizes endogenous levels of RIP protein only when phosphorylated at Ser166. This antibody is preferred for immunofluorescence whereas Phospho-RIP (Ser166) (D1L3S) Rabbit Monoclonal Antibody #65746 is preferred for western blot. Weak centriolar background staining was observed in some cell types.
Species Reactivity: Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser166 of human RIP 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). 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).
Alternate Names
AIEFL; Cell death protein RIP; FLJ39204; IMD57; receptor (TNFRSF)-interacting serine-threonine kinase 1; receptor interacting protein; receptor interacting serine/threonine kinase 1; Receptor-interacting protein 1; receptor-interacting protein kinase 1; Receptor-interacting serine/threonine-protein kinase 1; RIP; RIP-1; RIP1; RIPK1; Serine/threonine-protein kinase RIP
