CST, 32183S, Phospho-Glucocorticoid Receptor (Ser211) (F7L7X) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying Glucocorticoid Receptor (Ser211) phosphate. Validated for WB,IP,F,ChIP,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, Phospho-Glucocorticoid Receptor (Ser211) (F7L7X) Rabbit Monoclonal Antibody (CST #32183) is ready to ship. Product Usage Information For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. Western Blotting: 1:1000 Immunoprecipitation: 1:100 Flow Cytometry (Fixed/Permeabilized): 1:400 - 1:1600 Chromatin IP: 1:100 Chromatin IP-seq: 1:100 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. Protocol Available protocols: Western Blotting, Immunoprecipitation, Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq Specificity / Sensitivity Phospho-Glucocorticoid Receptor (Ser211) (F7L7X) Rabbit Monoclonal Antibody recognizes endogenous levels of glucocorticoid receptor protein only when phosphorylated at Ser211. This antibody detects a 250 kDa protein of unknown identity in some cell lines. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser211 of human glucocorticoid receptor protein. Background Glucocorticoid hormones control cellular proliferation, inflammation, and metabolism through their association with the glucocorticoid receptor (GR)/NR3C1, a member of the nuclear hormone receptor superfamily of transcription factors (1). GR is composed of several conserved structural elements, including a carboxy-terminal ligand-binding domain (which also contains residues critical for receptor dimerization and hormone-dependent gene transactivation), a neighboring hinge region containing nuclear localization signals, a central zinc-finger-containing DNA-binding domain, and an amino-terminal variable region that participates in ligand-independent gene transcription. In the absence of hormone, a significant population of GR is localized to the cytoplasm in an inactive form via its association with regulatory chaperone proteins, such as HSP90, HSP70, and FKBP52. On hormone binding, GR is released from the chaperone complex and translocates to the nucleus as a dimer to associate with specific DNA sequences termed glucocorticoid response elements (GREs), thereby enhancing or repressing transcription of specific target genes (2). It was demonstrated that GR-mediated transcriptional activation is modulated by phosphorylation (3-5). Although GR can be basally phosphorylated in the absence of hormone, it becomes hyperphosphorylated upon binding receptor agonists. It has been suggested that hormone-dependent phosphorylation of GR may determine target promoter specificity, cofactor interaction, strength and duration of receptor signaling, receptor stability, and receptor subcellular localization (3). Indeed, Ser211 of human GR is phosphorylated to a greater extent in the presence of hormone, and biochemical fractionation studies following hormone treatment indicate that Ser211-phosphorylated GR is found in the nucleus (3). Thus, Ser211 phosphorylation is a biomarker for activated GR . An added layer of complexity to GR signaling lies in the ability of multiple isoforms to be generated through both alternative splicing and the use of alternative translation initiation start sites, thus increasing the repertoire of functional signaling homo- and heterodimers (6,7). Alternate Names GCCR; GCR; GCRST; glucocorticoid nuclear receptor variant 1; Glucocorticoid receptor; GR; GRL; NR3C1; Nuclear receptor subfamily 3 group C member 1; nuclear receptor subfamily 3 group C member 1 variant hGR-B(54); nuclear receptor subfamily 3 group C member 1 variant hGR-B(77); nuclear receptor subfamily 3 group C member 1 variant hGR-B(93); nuclear receptor subfamily 3, group C, member 1 (glucocorticoid receptor) Specification REACTIVITY: H M R SENSITIVITY: Endogenous MW (kDa): 94, 91 Source/Isotype: Rabbit IgG