WB, IP, IF-IC, ChIP
H M
Endogenous
94, 91
Rabbit IgG
#P04150
2908
Product Information
Product Usage Information
For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:100 |
| Immunofluorescence (Immunocytochemistry) | 1:400 |
| Chromatin IP | 1:100 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser226 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).
Phosphorylation of GR at serine 226 by JNK enhances nuclear export after ligand depletion (6,7). Phosphorylation of various serine residues, including serine 226 also affect GR binding to different target genes, contributing to an additional layer of transcriptional regulation (8). Serine 226 phosphorylation has also been linked to depression disorders as well as inflammation (9-11).
- Yamamoto, K.R. (1985) Annu. Rev. Genet 19, 209-52.
- Necela, B.M. and Cidlowski, J.A. (2003) Trends Pharmacol. Sci. 24, 58-61.
- Wang, Z. et al. (2002) J. Biol. Chem. 277, 26573-80.
- Rogatsky, I. et al. (1998) J. Biol. Chem. 273, 14315-21.
- Krstic, M. D. et al. (1997) Mol. Cell. Biol. 17, 3947-54.
- Rogatsky, I. et al. (1998) Proc Natl Acad Sci U S A 95, 2050-5.
- Itoh, M. et al. (2002) Mol Endocrinol 16, 2382-92.
- Blind, R.D. and Garabedian, M.J. (2008) J Steroid Biochem Mol Biol 109, 150-7.
- Mitic, M. et al. (2013) Neuropharmacology 70, 100-11.
- Lukic, I. et al. (2015) J Mol Neurosci 55, 951-8.
- Verhoog, N.J. et al. (2011) J Biol Chem 286, 19297-310.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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