Immunofluorescent analysis of Glucocorticoid Receptor using Glucocorticoid Receptor Polyclonal Antibody (59) (Product# PA1-512 ) shows staining in A2058 Cells. Glucocorticoid Receptor (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucocorticoid Receptor (Product# PA1-512 ) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product# 35552 for GAR, Product# 35503 for GAM). Images were taken at 60X magnification.
|Tested species reactivity||Mouse, Rat, Human|
|Published species reactivity||Rat, Non-human primate, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C(245) K P L I L P D T K P K I K D(259) of human GR.|
|Purification||Antigen affinity chromatography|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||Assay dependent|
|Gel Shift (GS)||Assay dependent|
|Immunocytochemistry (ICC)||Assay dependent|
|Immunohistochemistry (Frozen) (IHC (F))||1 µg/ml|
|Immunoprecipitation (IP)||5 µg/ml|
|Western Blot (WB)||5 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-512 detects glucocorticoid receptor (GR) from human, mouse and rat tissues.
PA1-512 has been successfully used in Western blot, immunohistochemistry, immunocytochemistry, immunoprecipitation, immunofluorescence, ChIP and gel shift procedures. By Western blot, this antibody detects an ~97 kDa protein representing GR as well as two other unidentified proteins at ~135 to ~145 kDa from rat liver extract. These cross reacting proteins are not believed to be GR or GR precursors as they do not bind [3H]dexamethasone mesylate. Immunohistochemical staining of GR in rat hippocampus using PA1-512 results in staining primarily in the nucleus. PA1-512 does not disrupt receptor-DNA interactions.
The PA1-512 immunogen is a synthetic peptide corresponding to residues C(245) K P L I L P D T K P K I K D(259) of human GR. The PA1-512 immunizing peptide (Cat. # PEP-002) is available for use in for neutralization and control experiments.
Reconstitute with PBS.
Steroid receptors are ligand-dependent, intracellular proteins that stimulate transcription of specific genes by binding to specific DNA sequences following activation by the appropriate hormone. Glucocorticoids are a family of steroids necessary for the regulation of energy metabolism and the immune and inflammatory responses. These compounds exert their effect through their interaction with the glucocoticoid receptor (GR) and that complex's subsequent association with DNA. All normal mammalian tissues examined to date have been shown to contain glucocorticoid receptor.
The corresponding gene for the glucocorticoid receptor is NR3C1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Tumor suppressor protein (p)53, is a regulator of NF-kappaB repression by the glucocorticoid receptor.
PA1-512 was used in immunocytochemistry to investigate the effect of p53 on NF-kappaB repression by the glucocorticoid receptor
|Murphy SH,Suzuki K,Downes M,Welch GL,De Jesus P,Miraglia LJ,Orth AP,Chanda SK,Evans RM,Verma IM||Proceedings of the National Academy of Sciences of the United States of America (108:17117)||2011|
Functional modulation of the glucocorticoid receptor and suppression of NF-kappaB-dependent transcription by ursodeoxycholic acid.
PA1-512 was used in immunocytochemistry to study the molecular mechanism of UDCA action.
|Miura T,Ouchida R,Yoshikawa N,Okamoto K,Makino Y,Nakamura T,Morimoto C,Makino I,Tanaka H||The Journal of biological chemistry (276:47371)||2001|
Immunocytochemical localization of the glucocorticoid receptor in rat brain, pituitary, liver, and thymus with two new polyclonal antipeptide antibodies.
PA1-512 was used in immunocytochemistry to investigate the distribution of glucocorticoid receptor in various rat tissues
|McGimsey WC,Cidlowski JA,Stumpf WE,Sar M||Endocrinology (129:3064)||1991|
Progesterone receptor directly inhibits ß-casein gene transcription in mammary epithelial cells through promoting promoter and enhancer repressive chromatin modifications.
PA1-512 was used in ChIP assay to study the effect of progesterone receptor on casein gene transcription in mammary epithelial cells and its mechanism
|Buser AC,Obr AE,Kabotyanski EB,Grimm SL,Rosen JM,Edwards DP||Molecular endocrinology (Baltimore, Md.) (25:955)||2011|
Transcriptional regulation of the mouse IL-7 receptor alpha promoter by glucocorticoid receptor.
PA1-512 was used in chromatin immunoprecipitation to investigate the role of the glucocorticoid receptor during the transcriptional regulation of the mouse IL-7 receptor alpha promoter.
|Lee HC,Shibata H,Ogawa S,Maki K,Ikuta K||Journal of immunology (Baltimore, Md. : 1950) (174:7800)||2005|
STAT3-dependent enhanceosome assembly and disassembly: synergy with GR for full transcriptional increase of the alpha 2-macroglobulin gene.
PA1-512 was used in chromatin immunoprecipitation to investigate the role of GR during the STAT3-dependent enhanceosome assembly and disassembly.
|Lerner L,Henriksen MA,Zhang X,Darnell JE||Genes and development (17:2564)||2003|
Cross talk between glucocorticoid and estrogen receptors occurs at a subset of proinflammatory genes.
PA1-512 was used in western blot to study cross talk between glucocorticoid and estrogen receptors at a subset of proinflammatory genes
|Cvoro A,Yuan C,Paruthiyil S,Miller OH,Yamamoto KR,Leitman DC||Journal of immunology (Baltimore, Md. : 1950) (186:4354)||2011|
Bag-1M is a component of the in vivo DNA-glucocorticoid receptor complex at hormone-regulated promoter.
PA1-512 was used in western blot to investigate the influence of Bag-1M on gene expression modulated by glucocorticoid receptor
|Hong W,Baniahmad A,Liu Y,Li H||Journal of molecular biology (384:22)||2008|
Identification of a novel glucocorticoid receptor mutation in budesonide-resistant human bronchial epithelial cells.
PA1-512 was used in western blot to identify the mutation of a novel glucocorticoid receptor in budesonide-resistant human bronchial epithelial cells.
|Kunz S,Sandoval R,Carlsson P,Carlstedt-Duke J,Bloom JW,Miesfeld RL||Molecular endocrinology (Baltimore, Md.) (17:2566)||2003|
ATP-dependent mobilization of the glucocorticoid receptor during chromatin remodeling.
PA1-512 was used in western blot to investigate the mobilization movement of glucocorticoid receptor during chromatin remodeling process.
|Fletcher TM,Xiao N,Mautino G,Baumann CT,Wolford R,Warren BS,Hager GL||Molecular and cellular biology (22:3255)||2002|
Beta(2)-adrenergic receptors potentiate glucocorticoid receptor transactivation via G protein beta gamma-subunits and the phosphoinositide 3-kinase pathway.
PA1-512 was used in western blot to study the effect of beta(2)-adrenergic receptors on glucocorticoid receptor transactivation and its mechanism.
|Schmidt P,Holsboer F,Spengler D||Molecular endocrinology (Baltimore, Md.) (15:553)||2001|
Pharmacological characterization of glucocorticoid receptors in primary human bronchial epithelial cells.
PA1-512 was used in western blot to investigate the expression and activity of glucocorticoid receptors in primary bronchial epithelial cells
|LeVan TD,Babin EA,Yamamura HI,Bloom JW||Biochemical pharmacology (57:1003)||1999|
Glucocorticoid resistance in the squirrel monkey is associated with overexpression of the immunophilin FKBP51.
PA1-512 was used in western blot to study the expression of the hsp90-associated immunophilin FKBP51 in squirrel monkeys and its effect on glucocorticoid receptor.
|Reynolds PD,Ruan Y,Smith DF,Scammell JG||The Journal of clinical endocrinology and metabolism (84:663)||1999|
Transrepression of c-jun gene expression by the glucocorticoid receptor requires both AP-1 sites in the c-jun promoter.
PA1-512 was used in western blot to study the role of the glucocorticoid during the modulation of the c-jun promoter activity
|Wei P,Inamdar N,Vedeckis WV||Molecular endocrinology (Baltimore, Md.) (12:1322)||1998|
Restoration of the glucocorticoid receptor function by the phosphodiester compound of vitamins C and E, EPC-K1 (L-ascorbic acid 2-[3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1-benzopyran-6-yl hydrogen phosphate] potassium salt), via a redox-dependent mechanism.
PA1-512 was used in western blot to investigate the effect of redox state on glucocorticoid receptor function and the effect of antioxidants
|Okamoto K,Tanaka H,Makino Y,Makino I||Biochemical pharmacology (56:79)||1998|
Thioredoxin: a redox-regulating cellular cofactor for glucocorticoid hormone action. Cross talk between endocrine control of stress response and cellular antioxidant defense system.
PA1-512 was used in western blot to investigate the role of thioredoxin (TRX) on cellular sensitivity to glucocorticoids
|Makino Y,Okamoto K,Yoshikawa N,Aoshima M,Hirota K,Yodoi J,Umesono K,Makino I,Tanaka H||The Journal of clinical investigation (98:2469)||1996|
Charge heterogeneity in wildtype and variant glucocorticoid receptors.
PA1-512 was used in western blot to study the charge heterogeneity of glucocorticoid receptors from lymphosarcoma lines sensitive and resistant to glucocorticoid-mediated apoptosis
|Rowan BG,Ip MM||Molecular and cellular endocrinology (107:41)||1995|
Ligand-dependent down-regulation of stably transfected human glucocorticoid receptors is associated with the loss of functional glucocorticoid responsiveness.
PA1-512 was used in western blot to investigate the regulation of stably transfected human glucocorticoid receptors by glucocorticoids
|Bellingham DL,Sar M,Cidlowski JA||Molecular endocrinology (Baltimore, Md.) (6:2090)||1992|
Novel antipeptide antibodies to the human glucocorticoid receptor: recognition of multiple receptor forms in vitro and distinct localization of cytoplasmic and nuclear receptors.
PA1-512 was used in western blot to produce novel antibodies specifically recognizing human GR
|Cidlowski JA,Bellingham DL,Powell-Oliver FE,Lubahn DB,Sar M||Molecular endocrinology (Baltimore, Md.) (4:1427)||1990|
Human glucocorticoid receptor cDNA contains sequences sufficient for receptor down-regulation.
PA1-512 was used in western blot to investigate the intrinsic mechanism for the down-regulation of glucocorticoid receptor expression
|Burnstein KL,Jewell CM,Cidlowski JA||The Journal of biological chemistry (265:7284)||1990|
Stealth-nanoparticle strategy for enhancing the efficacy of steroids in mice with noise-induced hearing loss.
PA1-512 was used in immunohistochemistry to evaluate the therapeutic effect of nano-particles for steroid delivery in mice
|Horie RT,Sakamoto T,Nakagawa T,Ishihara T,Higaki M,Ito J||Nanomedicine (London, England) (5:1331)||2010|
The effect of long-term exposure to combinations of growth promoters in Long Evans rats: part 2. Adrenal morphology (histopathology and immunochemical studies).
PA1-512 was used in immunohistochemistry to investigate the effect of growth promoters on adrenal morphology and glucocorticoid receptor expression
|Illera JC,Peña L,Martínez-Mateos MM,Camacho L,Blass A,Garcia-Partida P,Illera MJ,Silván G||Analytica chimica acta (586:252)||2007|
Redox-dependent regulation of nuclear import of the glucocorticoid receptor.
PA1-512 was used in immunohistochemistry to investigate the role of the redox during the nuclear import of the glucocorticoid receptor
|Okamoto K,Tanaka H,Ogawa H,Makino Y,Eguchi H,Hayashi S,Yoshikawa N,Poellinger L,Umesono K,Makino I||The Journal of biological chemistry (274:10363)||1999|
Enhanced neutrophil expression of annexin-1 in coronary artery disease.
PA1-512 was used in flow cytometry to investigate the expression of annexin-1 in neutrophils in coronary artery disease patients
|Särndahl E,Bergström I,Nijm J,Forslund T,Perretti M,Jonasson L||Metabolism: clinical and experimental (59:433)||2010|
|Non-human primate||Not Cited||
ATP hydrolysis is essential for Bag-1M-mediated inhibition of the DNA binding by the glucocorticoid receptor.
PA1-512 was used in EMSA to study the involvement of ATP hydrolysis in the interplay of Bag-1M and glucocorticoid receptor
|Hong W,Chen L,Liu Y,Gao W||Biochemical and biophysical research communications (390:77)||2009|
|Non-human primate||Not Cited||
RAP46 is a negative regulator of glucocorticoid receptor action and hormone-induced apoptosis.
PA1-512 was used in EMSA assay to study the effect of RAP46 on glucocorticoid receptor action and hormone-induced apoptosis
|Kullmann M,Schneikert J,Moll J,Heck S,Zeiner M,Gehring U,Cato AC||The Journal of biological chemistry (273:14620)||1998|