PA1-512
antibody from Invitrogen Antibodies
Targeting: NR3C1
GR, GRL
Antibody data
- Antibody Data
- Antigen structure
- References [42]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [3]
- Other assay [2]
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Validation data
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- Product number
- PA1-512 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Glucocorticoid Receptor Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- 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 in 50 µL PBS to create a stock of 1 mg/mL.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 50 µg
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references GLCCI1 is a novel protector against glucocorticoid-induced apoptosis in T cells.
Corticosterone administration targeting a hypo-reactive HPA axis rescues a socially-avoidant phenotype in scarcity-adversity reared rats.
Tumor suppressor protein (p)53, is a regulator of NF-kappaB repression by the glucocorticoid receptor.
Progesterone receptor directly inhibits β-casein gene transcription in mammary epithelial cells through promoting promoter and enhancer repressive chromatin modifications.
Cross talk between glucocorticoid and estrogen receptors occurs at a subset of proinflammatory genes.
Stealth-nanoparticle strategy for enhancing the efficacy of steroids in mice with noise-induced hearing loss.
Enhanced neutrophil expression of annexin-1 in coronary artery disease.
Cortisol is involved in temperature-dependent sex determination in the Japanese flounder.
ATP hydrolysis is essential for Bag-1M-mediated inhibition of the DNA binding by the glucocorticoid receptor.
TH17 cells mediate steroid-resistant airway inflammation and airway hyperresponsiveness in mice.
Bag-1M is a component of the in vivo DNA-glucocorticoid receptor complex at hormone-regulated promoter.
A reduction state potentiates the glucocorticoid response through receptor protein stabilization.
The effect of long-term exposure to combinations of growth promoters in Long Evans rats: part 2. Adrenal morphology (histopathology and immunochemical studies).
A novel role for the glucocorticoid receptor in the regulation of monocyte chemoattractant protein-1 mRNA stability.
Integration of prolactin and glucocorticoid signaling at the beta-casein promoter and enhancer by ordered recruitment of specific transcription factors and chromatin modifiers.
Transcriptional regulation of the mouse IL-7 receptor alpha promoter by glucocorticoid receptor.
STAT3-dependent enhanceosome assembly and disassembly: synergy with GR for full transcriptional increase of the alpha 2-macroglobulin gene.
Identification of a novel glucocorticoid receptor mutation in budesonide-resistant human bronchial epithelial cells.
Role of the glucocorticoid receptor for regulation of hypoxia-dependent gene expression.
ATP-dependent mobilization of the glucocorticoid receptor during chromatin remodeling.
Functional modulation of the glucocorticoid receptor and suppression of NF-kappaB-dependent transcription by ursodeoxycholic acid.
Beta(2)-adrenergic receptors potentiate glucocorticoid receptor transactivation via G protein beta gamma-subunits and the phosphoinositide 3-kinase pathway.
Pharmacological characterization of glucocorticoid receptors in primary human bronchial epithelial cells.
Subcellular localization of glucocorticoid receptor protein in the human kidney glomerulus.
Glucocorticoid resistance in the squirrel monkey is associated with overexpression of the immunophilin FKBP51.
Redox-dependent regulation of nuclear import of the glucocorticoid receptor.
Redox-dependent regulation of nuclear import of the glucocorticoid receptor.
Transrepression of c-jun gene expression by the glucocorticoid receptor requires both AP-1 sites in the c-jun promoter.
RAP46 is a negative regulator of glucocorticoid receptor action and hormone-induced apoptosis.
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.
Expression and subcellular distribution of the beta-isoform of the human glucocorticoid receptor.
Coordinate regulation of glucocorticoid receptor and c-jun gene expression is cell type-specific and exhibits differential hormonal sensitivity for down- and up-regulation.
Ligand-independent activation of the glucocorticoid receptor by ursodeoxycholic acid. Repression of IFN-gamma-induced MHC class II gene expression via a glucocorticoid receptor-dependent pathway.
Thioredoxin: a redox-regulating cellular cofactor for glucocorticoid hormone action. Cross talk between endocrine control of stress response and cellular antioxidant defense system.
Zinc ions antagonize the inhibitory effect of aurothiomalate on glucocorticoid receptor function at physiological concentrations.
Charge heterogeneity in wildtype and variant glucocorticoid receptors.
A distinct modulating domain in glucocorticoid receptor monomers in the repression of activity of the transcription factor AP-1.
Ligand-dependent down-regulation of stably transfected human glucocorticoid receptors is associated with the loss of functional glucocorticoid responsiveness.
Evaluation of the role of ligand and thermal activation of specific DNA binding by in vitro synthesized human glucocorticoid receptor.
Immunocytochemical localization of the glucocorticoid receptor in rat brain, pituitary, liver, and thymus with two new polyclonal antipeptide antibodies.
Novel antipeptide antibodies to the human glucocorticoid receptor: recognition of multiple receptor forms in vitro and distinct localization of cytoplasmic and nuclear receptors.
Human glucocorticoid receptor cDNA contains sequences sufficient for receptor down-regulation.
Kiuchi Z, Nishibori Y, Kutsuna S, Kotani M, Hada I, Kimura T, Fukutomi T, Fukuhara D, Ito-Nitta N, Kudo A, Takata T, Ishigaki Y, Tomosugi N, Tanaka H, Matsushima S, Ogasawara S, Hirayama Y, Takematsu H, Yan K
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2019 Jun;33(6):7387-7402
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2019 Jun;33(6):7387-7402
Corticosterone administration targeting a hypo-reactive HPA axis rescues a socially-avoidant phenotype in scarcity-adversity reared rats.
Perry RE, Rincón-Cortés M, Braren SH, Brandes-Aitken AN, Opendak M, Pollonini G, Chopra D, Raver CC, Alberini CM, Blair C, Sullivan RM
Developmental cognitive neuroscience 2019 Dec;40:100716
Developmental cognitive neuroscience 2019 Dec;40:100716
Tumor suppressor protein (p)53, is a regulator of 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 2011 Oct 11;108(41):17117-22
Proceedings of the National Academy of Sciences of the United States of America 2011 Oct 11;108(41):17117-22
Progesterone receptor directly inhibits β-casein gene transcription in mammary epithelial cells through promoting promoter and enhancer repressive chromatin modifications.
Buser AC, Obr AE, Kabotyanski EB, Grimm SL, Rosen JM, Edwards DP
Molecular endocrinology (Baltimore, Md.) 2011 Jun;25(6):955-68
Molecular endocrinology (Baltimore, Md.) 2011 Jun;25(6):955-68
Cross talk between glucocorticoid and estrogen receptors occurs at a subset of proinflammatory genes.
Cvoro A, Yuan C, Paruthiyil S, Miller OH, Yamamoto KR, Leitman DC
Journal of immunology (Baltimore, Md. : 1950) 2011 Apr 1;186(7):4354-60
Journal of immunology (Baltimore, Md. : 1950) 2011 Apr 1;186(7):4354-60
Stealth-nanoparticle strategy for enhancing the efficacy of steroids in mice with noise-induced hearing loss.
Horie RT, Sakamoto T, Nakagawa T, Ishihara T, Higaki M, Ito J
Nanomedicine (London, England) 2010 Nov;5(9):1331-40
Nanomedicine (London, England) 2010 Nov;5(9):1331-40
Enhanced neutrophil expression of annexin-1 in coronary artery disease.
Särndahl E, Bergström I, Nijm J, Forslund T, Perretti M, Jonasson L
Metabolism: clinical and experimental 2010 Mar;59(3):433-40
Metabolism: clinical and experimental 2010 Mar;59(3):433-40
Cortisol is involved in temperature-dependent sex determination in the Japanese flounder.
Yamaguchi T, Yoshinaga N, Yazawa T, Gen K, Kitano T
Endocrinology 2010 Aug;151(8):3900-8
Endocrinology 2010 Aug;151(8):3900-8
ATP hydrolysis is essential for Bag-1M-mediated inhibition of the DNA binding by the glucocorticoid receptor.
Hong W, Chen L, Liu Y, Gao W
Biochemical and biophysical research communications 2009 Dec 4;390(1):77-81
Biochemical and biophysical research communications 2009 Dec 4;390(1):77-81
TH17 cells mediate steroid-resistant airway inflammation and airway hyperresponsiveness in mice.
McKinley L, Alcorn JF, Peterson A, Dupont RB, Kapadia S, Logar A, Henry A, Irvin CG, Piganelli JD, Ray A, Kolls JK
Journal of immunology (Baltimore, Md. : 1950) 2008 Sep 15;181(6):4089-97
Journal of immunology (Baltimore, Md. : 1950) 2008 Sep 15;181(6):4089-97
Bag-1M is a component of the in vivo DNA-glucocorticoid receptor complex at hormone-regulated promoter.
Hong W, Baniahmad A, Liu Y, Li H
Journal of molecular biology 2008 Dec 5;384(1):22-30
Journal of molecular biology 2008 Dec 5;384(1):22-30
A reduction state potentiates the glucocorticoid response through receptor protein stabilization.
Kitagawa H, Yamaoka I, Akimoto C, Kase I, Mezaki Y, Shimizu T, Kato S
Genes to cells : devoted to molecular & cellular mechanisms 2007 Nov;12(11):1281-7
Genes to cells : devoted to molecular & cellular mechanisms 2007 Nov;12(11):1281-7
The effect of long-term exposure to combinations of growth promoters in Long Evans rats: part 2. Adrenal morphology (histopathology and immunochemical studies).
Illera JC, Peña L, Martínez-Mateos MM, Camacho L, Blass A, Garcia-Partida P, Illera MJ, Silván G
Analytica chimica acta 2007 Mar 14;586(1-2):252-8
Analytica chimica acta 2007 Mar 14;586(1-2):252-8
A novel role for the glucocorticoid receptor in the regulation of monocyte chemoattractant protein-1 mRNA stability.
Dhawan L, Liu B, Blaxall BC, Taubman MB
The Journal of biological chemistry 2007 Apr 6;282(14):10146-52
The Journal of biological chemistry 2007 Apr 6;282(14):10146-52
Integration of prolactin and glucocorticoid signaling at the beta-casein promoter and enhancer by ordered recruitment of specific transcription factors and chromatin modifiers.
Kabotyanski EB, Huetter M, Xian W, Rijnkels M, Rosen JM
Molecular endocrinology (Baltimore, Md.) 2006 Oct;20(10):2355-68
Molecular endocrinology (Baltimore, Md.) 2006 Oct;20(10):2355-68
Transcriptional regulation of the mouse IL-7 receptor alpha promoter by glucocorticoid receptor.
Lee HC, Shibata H, Ogawa S, Maki K, Ikuta K
Journal of immunology (Baltimore, Md. : 1950) 2005 Jun 15;174(12):7800-6
Journal of immunology (Baltimore, Md. : 1950) 2005 Jun 15;174(12):7800-6
STAT3-dependent enhanceosome assembly and disassembly: synergy with GR for full transcriptional increase of the alpha 2-macroglobulin gene.
Lerner L, Henriksen MA, Zhang X, Darnell JE Jr
Genes & development 2003 Oct 15;17(20):2564-77
Genes & development 2003 Oct 15;17(20):2564-77
Identification of a novel glucocorticoid receptor mutation in budesonide-resistant human bronchial epithelial cells.
Kunz S, Sandoval R, Carlsson P, Carlstedt-Duke J, Bloom JW, Miesfeld RL
Molecular endocrinology (Baltimore, Md.) 2003 Dec;17(12):2566-82
Molecular endocrinology (Baltimore, Md.) 2003 Dec;17(12):2566-82
Role of the glucocorticoid receptor for regulation of hypoxia-dependent gene expression.
Kodama T, Shimizu N, Yoshikawa N, Makino Y, Ouchida R, Okamoto K, Hisada T, Nakamura H, Morimoto C, Tanaka H
The Journal of biological chemistry 2003 Aug 29;278(35):33384-91
The Journal of biological chemistry 2003 Aug 29;278(35):33384-91
ATP-dependent mobilization of the glucocorticoid receptor during chromatin remodeling.
Fletcher TM, Xiao N, Mautino G, Baumann CT, Wolford R, Warren BS, Hager GL
Molecular and cellular biology 2002 May;22(10):3255-63
Molecular and cellular biology 2002 May;22(10):3255-63
Functional modulation of the glucocorticoid receptor and suppression of NF-kappaB-dependent transcription by ursodeoxycholic acid.
Miura T, Ouchida R, Yoshikawa N, Okamoto K, Makino Y, Nakamura T, Morimoto C, Makino I, Tanaka H
The Journal of biological chemistry 2001 Dec 14;276(50):47371-8
The Journal of biological chemistry 2001 Dec 14;276(50):47371-8
Beta(2)-adrenergic receptors potentiate glucocorticoid receptor transactivation via G protein beta gamma-subunits and the phosphoinositide 3-kinase pathway.
Schmidt P, Holsboer F, Spengler D
Molecular endocrinology (Baltimore, Md.) 2001 Apr;15(4):553-64
Molecular endocrinology (Baltimore, Md.) 2001 Apr;15(4):553-64
Pharmacological characterization of glucocorticoid receptors in primary human bronchial epithelial cells.
LeVan TD, Babin EA, Yamamura HI, Bloom JW
Biochemical pharmacology 1999 May 1;57(9):1003-9
Biochemical pharmacology 1999 May 1;57(9):1003-9
Subcellular localization of glucocorticoid receptor protein in the human kidney glomerulus.
Yan K, Kudo A, Hirano H, Watanabe T, Tasaka T, Kataoka S, Nakajima N, Nishibori Y, Shibata T, Kohsaka T, Higashihara E, Tanaka H, Watanabe H, Nagasawa T, Awa S
Kidney international 1999 Jul;56(1):65-73
Kidney international 1999 Jul;56(1):65-73
Glucocorticoid resistance in the squirrel monkey is associated with overexpression of the immunophilin FKBP51.
Reynolds PD, Ruan Y, Smith DF, Scammell JG
The Journal of clinical endocrinology and metabolism 1999 Feb;84(2):663-9
The Journal of clinical endocrinology and metabolism 1999 Feb;84(2):663-9
Redox-dependent regulation of 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 1999 Apr 9;274(15):10363-71
The Journal of biological chemistry 1999 Apr 9;274(15):10363-71
Redox-dependent regulation of 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 1999 Apr 9;274(15):10363-71
The Journal of biological chemistry 1999 Apr 9;274(15):10363-71
Transrepression of c-jun gene expression by the glucocorticoid receptor requires both AP-1 sites in the c-jun promoter.
Wei P, Inamdar N, Vedeckis WV
Molecular endocrinology (Baltimore, Md.) 1998 Sep;12(9):1322-33
Molecular endocrinology (Baltimore, Md.) 1998 Sep;12(9):1322-33
RAP46 is a negative regulator of 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 1998 Jun 5;273(23):14620-5
The Journal of biological chemistry 1998 Jun 5;273(23):14620-5
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.
Okamoto K, Tanaka H, Makino Y, Makino I
Biochemical pharmacology 1998 Jul 1;56(1):79-86
Biochemical pharmacology 1998 Jul 1;56(1):79-86
Expression and subcellular distribution of the beta-isoform of the human glucocorticoid receptor.
Oakley RH, Webster JC, Sar M, Parker CR Jr, Cidlowski JA
Endocrinology 1997 Nov;138(11):5028-38
Endocrinology 1997 Nov;138(11):5028-38
Coordinate regulation of glucocorticoid receptor and c-jun gene expression is cell type-specific and exhibits differential hormonal sensitivity for down- and up-regulation.
Barrett TJ, Vig E, Vedeckis WV
Biochemistry 1996 Jul 30;35(30):9746-53
Biochemistry 1996 Jul 30;35(30):9746-53
Ligand-independent activation of the glucocorticoid receptor by ursodeoxycholic acid. Repression of IFN-gamma-induced MHC class II gene expression via a glucocorticoid receptor-dependent pathway.
Tanaka H, Makino Y, Miura T, Hirano F, Okamoto K, Komura K, Sato Y, Makino I
Journal of immunology (Baltimore, Md. : 1950) 1996 Feb 15;156(4):1601-8
Journal of immunology (Baltimore, Md. : 1950) 1996 Feb 15;156(4):1601-8
Thioredoxin: a redox-regulating cellular cofactor for glucocorticoid hormone action. Cross talk between endocrine control of stress response and cellular antioxidant defense system.
Makino Y, Okamoto K, Yoshikawa N, Aoshima M, Hirota K, Yodoi J, Umesono K, Makino I, Tanaka H
The Journal of clinical investigation 1996 Dec 1;98(11):2469-77
The Journal of clinical investigation 1996 Dec 1;98(11):2469-77
Zinc ions antagonize the inhibitory effect of aurothiomalate on glucocorticoid receptor function at physiological concentrations.
Tanaka H, Makino Y, Dahlman-Wright K, Gustafsson JA, Okamoto K, Makino I, Wright KD [corrected to Dahlman-Wright K]
Molecular pharmacology 1995 Nov;48(5):938-45
Molecular pharmacology 1995 Nov;48(5):938-45
Charge heterogeneity in wildtype and variant glucocorticoid receptors.
Rowan BG, Ip MM
Molecular and cellular endocrinology 1995 Jan;107(1):41-54
Molecular and cellular endocrinology 1995 Jan;107(1):41-54
A distinct modulating domain in glucocorticoid receptor monomers in the repression of activity of the transcription factor AP-1.
Heck S, Kullmann M, Gast A, Ponta H, Rahmsdorf HJ, Herrlich P, Cato AC
The EMBO journal 1994 Sep 1;13(17):4087-95
The EMBO journal 1994 Sep 1;13(17):4087-95
Ligand-dependent down-regulation of stably transfected human glucocorticoid receptors is associated with the loss of functional glucocorticoid responsiveness.
Bellingham DL, Sar M, Cidlowski JA
Molecular endocrinology (Baltimore, Md.) 1992 Dec;6(12):2090-102
Molecular endocrinology (Baltimore, Md.) 1992 Dec;6(12):2090-102
Evaluation of the role of ligand and thermal activation of specific DNA binding by in vitro synthesized human glucocorticoid receptor.
Burnstein KL, Jewell CM, Cidlowski JA
Molecular endocrinology (Baltimore, Md.) 1991 Jul;5(7):1013-22
Molecular endocrinology (Baltimore, Md.) 1991 Jul;5(7):1013-22
Immunocytochemical localization of the glucocorticoid receptor in rat brain, pituitary, liver, and thymus with two new polyclonal antipeptide antibodies.
McGimsey WC, Cidlowski JA, Stumpf WE, Sar M
Endocrinology 1991 Dec;129(6):3064-72
Endocrinology 1991 Dec;129(6):3064-72
Novel antipeptide antibodies to the human glucocorticoid receptor: recognition of multiple receptor forms in vitro and distinct localization of cytoplasmic and nuclear receptors.
Cidlowski JA, Bellingham DL, Powell-Oliver FE, Lubahn DB, Sar M
Molecular endocrinology (Baltimore, Md.) 1990 Oct;4(10):1427-37
Molecular endocrinology (Baltimore, Md.) 1990 Oct;4(10):1427-37
Human glucocorticoid receptor cDNA contains sequences sufficient for receptor down-regulation.
Burnstein KL, Jewell CM, Cidlowski JA
The Journal of biological chemistry 1990 May 5;265(13):7284-91
The Journal of biological chemistry 1990 May 5;265(13):7284-91
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Glucocorticoid Receptor was performed by loading 20 µg of the indicated whole cell lysates and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. Glucocorticoid Receptor was detected at ~95 kDa using a Glucocorticoid Receptor rabbit polyclonal antibody (Product # PA1-512) at a concentration of 1 µg/mL in blocking buffer overnight at 4C on a rocking platform, followed by a Goat anti-rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) at a dilution of 1:1000 for at least one hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Glucocorticoid Receptor was achieved by transfecting MDA-MB-231 with Glucocorticoid Receptor specific siRNAs (Silencer® select Product # s6186 and s6187). Western blot analysis (Fig. a) was performed using whole cell extracts from the Glucocorticoid Receptor knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Glucocorticoid Receptor Polyclonal Antibody (Product # PA1-512, 0.15 µg/ml) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Glucocorticoid Receptor.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Glucocorticoid Receptor Polyclonal Antibody (Product # PA1-512) and a 86 kDa band corresponding to Glucocorticoid Receptor was observed across cell lines and tissues. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MDA-MB-231 (Lane 2), MCF7 (Lane 3), DU 145 (Lane 4) in Figure a and Mouse Skeletal Muscle (Lane 1), Mouse Brain (Lane 2), Rat Skeletal Muscle (Lane 3) and Rat Heart (Lane 4) in Figure b, were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.75 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Uncharacterized bands (*) were observed at 38 kDa, 55 kDa and 180 kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Glucocorticoid Receptor using Glucocorticoid Receptor Polyclonal Antibody (59) (Product # PA1-512) shows staining in Hela 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 1:100 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Glucocorticoid Receptor was performed using MDA-MB-231 cells (serum-starved) and MDA-MB-231 cells serum-starved for 24 hours, followed by 1 µM Dexamethasone treatment for 2 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Glucocorticoid Receptor Polyclonal Antibody (Product # PA1-512) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green) in MDA-MB-231 treated cells. Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization of NR3C1 protein in MDA-MB-231 treated cells. Panel e represents the merged image of MDA-MB-231 untreated cells, that shows cytoplasmic localization of NR3C1 protein. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL