PA1-516

antibody from Invitrogen Antibodies

Targeting: NR3C1 GR, GRL

Provider product page for PA1-516

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Chromatin Immunoprecipitation

Other assay

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

NR3C1 protein structure - PA1-516 shown in red.

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.

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 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.

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 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.

Supportive validation

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.

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.