PA1-516
antibody from Invitrogen Antibodies
Targeting: NR3C1
GR, GRL
Antibody data
- Antibody Data
- Antigen structure
- References [43]
- Comments [0]
- Validations
- Immunocytochemistry [3]
- Immunohistochemistry [3]
- Other assay [2]
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- Product number
- PA1-516 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Glucocorticoid Receptor alpha Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- PA1-516 detects glucocorticoid receptor (GR) alpha from human, mouse and rat tissues. This antibody does not detect GR beta. PA1-516 has been successfully used in Western blot, immunocytochemistry, immunofluorescence and immunoprecipitation (ChIP) procedures. By Western blot, this antibody detects an ~94 kDa protein representing GR in HeLa cell lysate. PA1-516 immunizing peptide corresponds to amino acid residues 755-771 of human GR alpha. This peptide (Cat. # PEP-221) is available for use in neutralization and control experiments. Reconstitute with 50 µL of distilled water.
- 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 Spatiotemporal control of axillary meristem formation by interacting transcriptional regulators.
Coordination of meristem and boundary functions by transcription factors in the SHOOT MERISTEMLESS regulatory network.
The Role of S-Palmitoylation of the Human Glucocorticoid Receptor (hGR) in Mediating the Nongenomic Glucocorticoid Actions.
Annexin A1 in blood mononuclear cells from patients with coronary artery disease: Its association with inflammatory status and glucocorticoid sensitivity.
The bHLH transcription factor SPATULA enables cytokinin signaling, and both activate auxin biosynthesis and transport genes at the medial domain of the gynoecium.
Two-Step Regulation of a Meristematic Cell Population Acting in Shoot Branching in Arabidopsis.
Additive anti-inflammatory effects of corticosteroids and phosphodiesterase-4 inhibitors in COPD CD8 cells.
Three Novel Heterozygous Point Mutations of NR3C1 Causing Glucocorticoid Resistance.
Dynamics of chromatin accessibility and gene regulation by MADS-domain transcription factors in flower development.
A kinase-independent activity of Cdk9 modulates glucocorticoid receptor-mediated gene induction.
Effect of ionizing radiation on human skeletal muscle precursor cells.
Glucocorticoid signaling in myeloid cells worsens acute CNS injury and inflammation.
The cholinergic and non-cholinergic effects of organophosphates and oximes in cultured human myoblasts.
PA1 protein, a new competitive decelerator acting at more than one step to impede glucocorticoid receptor-mediated transactivation.
Identification of location and kinetically defined mechanism of cofactors and reporter genes in the cascade of steroid-regulated transactivation.
Expression of glucocorticoid receptors in the regenerating human skeletal muscle.
Expression of glucocorticoid receptors in the regenerating human skeletal muscle.
Dynamic molecular and anatomical changes in the glucocorticoid receptor in human cortical development.
Abnormal glucocorticoid receptor mRNA and protein isoform expression in the prefrontal cortex in psychiatric illness.
Enhanced neutrophil expression of annexin-1 in coronary artery disease.
HIF-1alpha response to hypoxia is functionally separated from the glucocorticoid stress response in the in vitro regenerating human skeletal muscle.
Glucocorticoids with different chemical structures but similar glucocorticoid receptor potency regulate subsets of common and unique genes in human trabecular meshwork cells.
Potential role of glucocorticoid signaling in the formation of pancreatic islets in the human fetus.
Mutations of glucocorticoid receptor differentially affect AF2 domain activity in a steroid-selective manner to alter the potency and efficacy of gene induction and repression.
ATF2 impairs glucocorticoid receptor-mediated transactivation in human CD8+ T cells.
Participation of the Arabidopsis bHLH factor GL3 in trichome initiation regulatory events.
STAMP, a novel predicted factor assisting TIF2 actions in glucocorticoid receptor-mediated induction and repression.
Distribution of glucocorticoid receptor alpha and beta subtypes in the idiopathic inflammatory myopathies.
CD38 expression is insensitive to steroid action in cells treated with tumor necrosis factor-alpha and interferon-gamma by a mechanism involving the up-regulation of the glucocorticoid receptor beta isoform.
Increased glucocorticoid receptor and 11{beta}-hydroxysteroid dehydrogenase type 1 expression in hepatocytes may contribute to the phenotype of type 2 diabetes in db/db mice.
Glucocorticoid receptor subunit expression in adenotonsillar tissue of children with obstructive sleep apnea.
The Smad6-histone deacetylase 3 complex silences the transcriptional activity of the glucocorticoid receptor: potential clinical implications.
Superantigen-induced corticosteroid resistance of human T cells occurs through activation of the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK-ERK) pathway.
CD56+ cells induce steroid resistance in B cells exposed to IL-15.
Natural glucocorticoid receptor mutants causing generalized glucocorticoid resistance: molecular genotype, genetic transmission, and clinical phenotype.
Steroid-responsive sequences in the human glucocorticoid receptor gene 1A promoter.
The glucocorticoid receptor and the orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II interact with and mutually affect each other's transcriptional activities: implications for intermediary metabolism.
Effect of labor on glucocorticoid receptor (GR(Total), GRalpha, and GRbeta) proteins in ovine intrauterine tissues.
Effect of labor on glucocorticoid receptor (GR(Total), GRalpha, and GRbeta) proteins in ovine intrauterine tissues.
High constitutive glucocorticoid receptor beta in human neutrophils enables them to reduce their spontaneous rate of cell death in response to corticosteroids.
GRKO mice express an aberrant dexamethasone-binding glucocorticoid receptor, but are profoundly glucocorticoid resistant.
Decreased steroid responsiveness at night in nocturnal asthma. Is the macrophage responsible?
The human glucocorticoid receptor beta isoform. Expression, biochemical properties, and putative function.
Zhang C, Wang J, Wenkel S, Chandler JW, Werr W, Jiao Y
Development (Cambridge, England) 2018 Dec 10;145(24)
Development (Cambridge, England) 2018 Dec 10;145(24)
Coordination of meristem and boundary functions by transcription factors in the SHOOT MERISTEMLESS regulatory network.
Scofield S, Murison A, Jones A, Fozard J, Aida M, Band LR, Bennett M, Murray JAH
Development (Cambridge, England) 2018 Apr 30;145(9)
Development (Cambridge, England) 2018 Apr 30;145(9)
The Role of S-Palmitoylation of the Human Glucocorticoid Receptor (hGR) in Mediating the Nongenomic Glucocorticoid Actions.
Nicolaides NC, Kino T, Roberts ML, Katsantoni E, Sertedaki A, Moutsatsou P, Psarra AG, Chrousos GP, Charmandari E
Journal of molecular biochemistry 2017;6(1):3-12
Journal of molecular biochemistry 2017;6(1):3-12
Annexin A1 in blood mononuclear cells from patients with coronary artery disease: Its association with inflammatory status and glucocorticoid sensitivity.
Bergström I, Lundberg AK, Jönsson S, Särndahl E, Ernerudh J, Jonasson L
PloS one 2017;12(3):e0174177
PloS one 2017;12(3):e0174177
The bHLH transcription factor SPATULA enables cytokinin signaling, and both activate auxin biosynthesis and transport genes at the medial domain of the gynoecium.
Reyes-Olalde JI, Zúñiga-Mayo VM, Serwatowska J, Chavez Montes RA, Lozano-Sotomayor P, Herrera-Ubaldo H, Gonzalez-Aguilera KL, Ballester P, Ripoll JJ, Ezquer I, Paolo D, Heyl A, Colombo L, Yanofsky MF, Ferrandiz C, Marsch-Martínez N, de Folter S
PLoS genetics 2017 Apr;13(4):e1006726
PLoS genetics 2017 Apr;13(4):e1006726
Two-Step Regulation of a Meristematic Cell Population Acting in Shoot Branching in Arabidopsis.
Shi B, Zhang C, Tian C, Wang J, Wang Q, Xu T, Xu Y, Ohno C, Sablowski R, Heisler MG, Theres K, Wang Y, Jiao Y
PLoS genetics 2016 Jul;12(7):e1006168
PLoS genetics 2016 Jul;12(7):e1006168
Additive anti-inflammatory effects of corticosteroids and phosphodiesterase-4 inhibitors in COPD CD8 cells.
Grundy S, Plumb J, Kaur M, Ray D, Singh D
Respiratory research 2016 Jan 25;17:9
Respiratory research 2016 Jan 25;17:9
Three Novel Heterozygous Point Mutations of NR3C1 Causing Glucocorticoid Resistance.
Vitellius G, Fagart J, Delemer B, Amazit L, Ramos N, Bouligand J, Le Billan F, Castinetti F, Guiochon-Mantel A, Trabado S, Lombès M
Human mutation 2016 Aug;37(8):794-803
Human mutation 2016 Aug;37(8):794-803
Dynamics of chromatin accessibility and gene regulation by MADS-domain transcription factors in flower development.
Pajoro A, Madrigal P, Muiño JM, Matus JT, Jin J, Mecchia MA, Debernardi JM, Palatnik JF, Balazadeh S, Arif M, Ó'Maoiléidigh DS, Wellmer F, Krajewski P, Riechmann JL, Angenent GC, Kaufmann K
Genome biology 2014 Mar 3;15(3):R41
Genome biology 2014 Mar 3;15(3):R41
A kinase-independent activity of Cdk9 modulates glucocorticoid receptor-mediated gene induction.
Zhu R, Lu X, Pradhan M, Armstrong SP, Storchan GB, Chow CC, Simons SS Jr
Biochemistry 2014 Mar 25;53(11):1753-67
Biochemistry 2014 Mar 25;53(11):1753-67
Effect of ionizing radiation on human skeletal muscle precursor cells.
Jurdana M, Cemazar M, Pegan K, Mars T
Radiology and oncology 2013;47(4):376-81
Radiology and oncology 2013;47(4):376-81
Glucocorticoid signaling in myeloid cells worsens acute CNS injury and inflammation.
Sorrells SF, Caso JR, Munhoz CD, Hu CK, Tran KV, Miguel ZD, Chien BY, Sapolsky RM
The Journal of neuroscience : the official journal of the Society for Neuroscience 2013 May 1;33(18):7877-89
The Journal of neuroscience : the official journal of the Society for Neuroscience 2013 May 1;33(18):7877-89
The cholinergic and non-cholinergic effects of organophosphates and oximes in cultured human myoblasts.
Katalinić M, Miš K, Pirkmajer S, Grubič Z, Kovarik Z, Marš T
Chemico-biological interactions 2013 Mar 25;203(1):144-8
Chemico-biological interactions 2013 Mar 25;203(1):144-8
PA1 protein, a new competitive decelerator acting at more than one step to impede glucocorticoid receptor-mediated transactivation.
Zhang Z, Sun Y, Cho YW, Chow CC, Simons SS Jr
The Journal of biological chemistry 2013 Jan 4;288(1):42-58
The Journal of biological chemistry 2013 Jan 4;288(1):42-58
Identification of location and kinetically defined mechanism of cofactors and reporter genes in the cascade of steroid-regulated transactivation.
Blackford JA Jr, Guo C, Zhu R, Dougherty EJ, Chow CC, Simons SS Jr
The Journal of biological chemistry 2012 Nov 30;287(49):40982-95
The Journal of biological chemistry 2012 Nov 30;287(49):40982-95
Expression of glucocorticoid receptors in the regenerating human skeletal muscle.
Filipović D, Pirkmajer S, Mis K, Mars T, Grubic Z
Physiological research 2011;60(Suppl 1):S147-54
Physiological research 2011;60(Suppl 1):S147-54
Expression of glucocorticoid receptors in the regenerating human skeletal muscle.
Filipović D, Pirkmajer S, Mis K, Mars T, Grubic Z
Physiological research 2011;60 Suppl 1:S147-54
Physiological research 2011;60 Suppl 1:S147-54
Dynamic molecular and anatomical changes in the glucocorticoid receptor in human cortical development.
Sinclair D, Webster MJ, Wong J, Weickert CS
Molecular psychiatry 2011 May;16(5):504-15
Molecular psychiatry 2011 May;16(5):504-15
Abnormal glucocorticoid receptor mRNA and protein isoform expression in the prefrontal cortex in psychiatric illness.
Sinclair D, Tsai SY, Woon HG, Weickert CS
Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2011 Dec;36(13):2698-709
Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2011 Dec;36(13):2698-709
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
HIF-1alpha response to hypoxia is functionally separated from the glucocorticoid stress response in the in vitro regenerating human skeletal muscle.
Pirkmajer S, Filipovic D, Mars T, Mis K, Grubic Z
American journal of physiology. Regulatory, integrative and comparative physiology 2010 Dec;299(6):R1693-700
American journal of physiology. Regulatory, integrative and comparative physiology 2010 Dec;299(6):R1693-700
Glucocorticoids with different chemical structures but similar glucocorticoid receptor potency regulate subsets of common and unique genes in human trabecular meshwork cells.
Nehmé A, Lobenhofer EK, Stamer WD, Edelman JL
BMC medical genomics 2009 Sep 10;2:58
BMC medical genomics 2009 Sep 10;2:58
Potential role of glucocorticoid signaling in the formation of pancreatic islets in the human fetus.
Phan-Hug F, Guimiot F, Lelièvre V, Delezoide AL, Czernichow P, Breant B, Blondeau B
Pediatric research 2008 Oct;64(4):346-51
Pediatric research 2008 Oct;64(4):346-51
Mutations of glucocorticoid receptor differentially affect AF2 domain activity in a steroid-selective manner to alter the potency and efficacy of gene induction and repression.
Tao YG, Xu Y, Xu HE, Simons SS Jr
Biochemistry 2008 Jul 22;47(29):7648-62
Biochemistry 2008 Jul 22;47(29):7648-62
ATF2 impairs glucocorticoid receptor-mediated transactivation in human CD8+ T cells.
Li LB, Leung DY, Strand MJ, Goleva E
Blood 2007 Sep 1;110(5):1570-7
Blood 2007 Sep 1;110(5):1570-7
Participation of the Arabidopsis bHLH factor GL3 in trichome initiation regulatory events.
Morohashi K, Zhao M, Yang M, Read B, Lloyd A, Lamb R, Grotewold E
Plant physiology 2007 Nov;145(3):736-46
Plant physiology 2007 Nov;145(3):736-46
STAMP, a novel predicted factor assisting TIF2 actions in glucocorticoid receptor-mediated induction and repression.
He Y, Simons SS Jr
Molecular and cellular biology 2007 Feb;27(4):1467-85
Molecular and cellular biology 2007 Feb;27(4):1467-85
Distribution of glucocorticoid receptor alpha and beta subtypes in the idiopathic inflammatory myopathies.
De Bleecker JL, De Paepe B, Vervaet VL, Arys B, Creus KK, Werbrouck BF, Martin JJ
Neuromuscular disorders : NMD 2007 Feb;17(2):186-93
Neuromuscular disorders : NMD 2007 Feb;17(2):186-93
CD38 expression is insensitive to steroid action in cells treated with tumor necrosis factor-alpha and interferon-gamma by a mechanism involving the up-regulation of the glucocorticoid receptor beta isoform.
Tliba O, Cidlowski JA, Amrani Y
Molecular pharmacology 2006 Feb;69(2):588-96
Molecular pharmacology 2006 Feb;69(2):588-96
Increased glucocorticoid receptor and 11{beta}-hydroxysteroid dehydrogenase type 1 expression in hepatocytes may contribute to the phenotype of type 2 diabetes in db/db mice.
Liu Y, Nakagawa Y, Wang Y, Sakurai R, Tripathi PV, Lutfy K, Friedman TC
Diabetes 2005 Jan;54(1):32-40
Diabetes 2005 Jan;54(1):32-40
Glucocorticoid receptor subunit expression in adenotonsillar tissue of children with obstructive sleep apnea.
Goldbart AD, Veling MC, Goldman JL, Li RC, Brittian KR, Gozal D
Pediatric research 2005 Feb;57(2):232-6
Pediatric research 2005 Feb;57(2):232-6
The Smad6-histone deacetylase 3 complex silences the transcriptional activity of the glucocorticoid receptor: potential clinical implications.
Ichijo T, Voutetakis A, Cotrim AP, Bhattachryya N, Fujii M, Chrousos GP, Kino T
The Journal of biological chemistry 2005 Dec 23;280(51):42067-77
The Journal of biological chemistry 2005 Dec 23;280(51):42067-77
Superantigen-induced corticosteroid resistance of human T cells occurs through activation of the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK-ERK) pathway.
Li LB, Goleva E, Hall CF, Ou LS, Leung DY
The Journal of allergy and clinical immunology 2004 Nov;114(5):1059-69
The Journal of allergy and clinical immunology 2004 Nov;114(5):1059-69
CD56+ cells induce steroid resistance in B cells exposed to IL-15.
Xu Q, Goleva E, Ou LS, Li LB, Leung DY
Journal of immunology (Baltimore, Md. : 1950) 2004 Jun 1;172(11):7110-5
Journal of immunology (Baltimore, Md. : 1950) 2004 Jun 1;172(11):7110-5
Natural glucocorticoid receptor mutants causing generalized glucocorticoid resistance: molecular genotype, genetic transmission, and clinical phenotype.
Charmandari E, Kino T, Souvatzoglou E, Vottero A, Bhattacharyya N, Chrousos GP
The Journal of clinical endocrinology and metabolism 2004 Apr;89(4):1939-49
The Journal of clinical endocrinology and metabolism 2004 Apr;89(4):1939-49
Steroid-responsive sequences in the human glucocorticoid receptor gene 1A promoter.
Geng CD, Vedeckis WV
Molecular endocrinology (Baltimore, Md.) 2004 Apr;18(4):912-24
Molecular endocrinology (Baltimore, Md.) 2004 Apr;18(4):912-24
The glucocorticoid receptor and the orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II interact with and mutually affect each other's transcriptional activities: implications for intermediary metabolism.
De Martino MU, Bhattachryya N, Alesci S, Ichijo T, Chrousos GP, Kino T
Molecular endocrinology (Baltimore, Md.) 2004 Apr;18(4):820-33
Molecular endocrinology (Baltimore, Md.) 2004 Apr;18(4):820-33
Effect of labor on glucocorticoid receptor (GR(Total), GRalpha, and GRbeta) proteins in ovine intrauterine tissues.
Gupta S, Gyomorey S, Lye SJ, Gibb W, Challis JR
Journal of the Society for Gynecologic Investigation 2003 Apr;10(3):136-44
Journal of the Society for Gynecologic Investigation 2003 Apr;10(3):136-44
Effect of labor on glucocorticoid receptor (GR(Total), GRalpha, and GRbeta) proteins in ovine intrauterine tissues.
Gupta S, Gyomorey S, Lye SJ, Gibb W, Challis JR
Journal of the Society for Gynecologic Investigation 2003 Apr;10(3):136-44
Journal of the Society for Gynecologic Investigation 2003 Apr;10(3):136-44
High constitutive glucocorticoid receptor beta in human neutrophils enables them to reduce their spontaneous rate of cell death in response to corticosteroids.
Strickland I, Kisich K, Hauk PJ, Vottero A, Chrousos GP, Klemm DJ, Leung DY
The Journal of experimental medicine 2001 Mar 5;193(5):585-93
The Journal of experimental medicine 2001 Mar 5;193(5):585-93
GRKO mice express an aberrant dexamethasone-binding glucocorticoid receptor, but are profoundly glucocorticoid resistant.
Cole TJ, Myles K, Purton JF, Brereton PS, Solomon NM, Godfrey DI, Funder JW
Molecular and cellular endocrinology 2001 Feb 28;173(1-2):193-202
Molecular and cellular endocrinology 2001 Feb 28;173(1-2):193-202
Decreased steroid responsiveness at night in nocturnal asthma. Is the macrophage responsible?
Kraft M, Hamid Q, Chrousos GP, Martin RJ, Leung DY
American journal of respiratory and critical care medicine 2001 Apr;163(5):1219-25
American journal of respiratory and critical care medicine 2001 Apr;163(5):1219-25
The human glucocorticoid receptor beta isoform. Expression, biochemical properties, and putative function.
Oakley RH, Sar M, Cidlowski JA
The Journal of biological chemistry 1996 Apr 19;271(16):9550-9
The Journal of biological chemistry 1996 Apr 19;271(16):9550-9
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Glucocorticoid Receptor alpha using Glucocorticoid Receptor alpha Polyclonal Antibody (Product # PA1-516) shows staining in A2058 Cells. Glucocorticoid Receptor alpha (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 alpha (Product # PA1-516) at a dilution of 1:200 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 alpha using Glucocorticoid Receptor alpha Polyclonal Antibody (Product # PA1-516) shows staining in Hela Cells. Glucocorticoid Receptor alpha (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 alpha (Product # PA1-516) 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
- Immunofluorescent analysis of Glucocorticoid Receptor alpha using Glucocorticoid Receptor alpha Polyclonal Antibody (Product # PA1-516) shows staining in U251 Cells. Glucocorticoid Receptor alpha (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 alpha (Product # PA1-516) 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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on cancer biopsies of deparaffinized Human cervical carcinoma tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha (Product # PA1-516) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on normal deparaffinized Human heart tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha (Product # PA1-516) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on normal deparaffinized Human tonsil tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha (Product # PA1-516) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 4 Effect of Dexamethasone and PDE4 inhibitors on nuclear translocation of Glucocorticoid Receptor. Peripheral blood CD8 cells from COPD patients ( n = 5) were untreated (basal), treated with GSK256066 (10 -9 M), roflumilast (10 -7 M), or forskolin (10 -6 M) alone or treated with dexamethasone (10 -6 M, Dex) alone and in combination with GSK256066, roflumilast or forskolin. Cells were harvested after 60 min. Cells were immunostained with rabbit anti-glucocorticoid receptor (GR) antibody (red). Nuclei were counterstained with 4',6-diamidino-2-phenylindole (blue). The nuclear mean fluorescence intensity (MFI) of GR was measured. Representative images of each experimental treatment are shown ( a - g ). h Data is presented as mean +- SE GR nuclear MFI normalised to basal levels. Statistically significant increase in nuclear GR above basal is indicated when * p < 0.05
- Submitted by
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- Figure 3. Identification of GL2 , CPC , and ETC1 as GL3 immediate direct targets. A, qRT-PCR of mRNA obtained from green tissues of 10-d-old gl3 egl3 plants expressing p35S::GL3-GR treated with DEX (black), or DEX and CHX (hatched) for 4 h. The bars indicate the relative expression to mock- or CHX-treated plants, respectively. Error bars indicate the sd of the data obtained from three independent RT-PCR reactions obtained from one induction experiment. All experiments were conducted in at least three biological replicates (three independent inductions), which showed no significant difference with each other. B, Semiquantitative PCR of ChIP experiments of gl3 egl3 p35S::GL3-GR plants mock-treated (M) or treated with DEX (D) for 4 h. Input corresponds to the chromatin prior to immunoprecipitation, alpha GR to the material recovered after immunoprecipitation with antibodies against GR. PCR was performed on three 4-fold serial dilutions of the ChIP-ed material, represented by the black slope on the top. The graph on the left indicates the position of the PCR fragment relative to the transcription start site (indicated by an arrow) of the various genes. At3g33520 and ACT2/7 correspond to negative controls.