Phospho-Glucocorticoid Recepto

4161:

Immunofluorescence , Immunoprecipitation , Western Blotting

Phospho-Glucocorticoid Receptor (Ser211) Antibody detects endogenous levels of glucocorticoid receptor only when phosphorylated at serine 211. This antibody does not cross-react with other unrelated phosphorylated proteins.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding serine 211 of human glucocorticoid receptor. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from A549(CCL-185) cells, untreated or stimulated with dexamethasone (100 nM for 1 hr), using Phospho-Glucocorticoid Receptor (Ser211) Antibody (upper) or control glucocorticoid receptor antibody (lower).

IF-IC

Confocal immunofluorescent analysis of HeLa cells, dexamethasone-treated (left) or lambda phosphatase-treated (right) using Phospho-Glucocorticoid Receptor (Ser211) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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 COOH-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 NH2-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 cell nucleus as a dimer to associate with specific DNA sequences termed glucocorticoid response elements (GREs), and increases or represses 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 in vivo . An added layer of complexity to GR signaling lies in the ability of mutiple isoforms to be generated through both alternative splicing and the use of alternative translation intiation start sites, thus increasing the repertoire of functional signaling homo- and heterodimers (6,7).

1. Yamamoto, K.R. (1985) Annu. Rev. Genet 19, 209-252.
2. Necela, B.M. and Cidlowski, J.A. (2003) Trends Pharmacol. Sci. 24, 58-61.
3. Wang, Z. et al. (2002) J. Biol. Chem. 277, 26573-26580.
4. Rogatsky, I. et al. (1998) J. Biol. Chem. 273, 14315-14321.
5. Krstic, M. D. et al. (1997) Mol. Cell. Biol. 17, 3947-3954.
6. Yudt, M.R. and Cidlowski, J.A. (2001) Mol Endocrinol 15, 1093-103.
7. Lu, N.Z. and Cidlowski, J.A. (2005) Mol Cell 18, 331-42.

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• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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