Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] - BSA and Azide Free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] - BSA and Azide Free (AB302678)

This data was developed using ab302677, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human prostatic hyperplasia tissue labeling Glucocorticoid receptor with ab302677 at 1/500 dilution (1.798 µg/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human prostatic hyperplasia is observed. The section was incubated with ab302677 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] - BSA and Azide Free (AB302678)

This data was developed using ab302677, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue labeling Glucocorticoid receptor with ab302677 at 1/500 dilution (1.798 µg/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human cervical cancer is observed. The section was incubated with ab302677 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] - BSA and Azide Free (AB302678)

This data was developed using ab302677, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Glucocorticoid receptor with ab302677 at 1/50 dilution (17.98 µg/mL), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing all the signal translocated from cytoplasm into nucleus in HeLa cells treated with dexamethasone (1 µM) for 20 min. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/50 dilution (10 µg/mL) (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL).

• WB

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Western blot - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] - BSA and Azide Free (AB302678)

This data was developed using ab302677, the same antibody clone in a different buffer formulation.

ab302677 was shown to react with NR3C1 in wild-type A549 cells in Western blot with loss of signal observed in a NR3C1 knockout cell line. Wild-type A549 and NR3C1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab302677 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] (<a href='/en-us/products/primary-antibodies/glucocorticoid-receptor-antibody-41-glucocorticoid-receptor-ab302677'>ab302677</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 lysate at 30 µg

Lane 2:

NR3C1 knock-out A549 lysate at 30 µg

Observed band size: 94 kDa,67 kDa

false

• WB

Supplier Data

Western blot - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] - BSA and Azide Free (AB302678)

This data was developed using ab302677, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS. Lysates at 20 µg per lane. False colour image of Western blot : Anti- Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] (ab302677 staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody 16891 loading control staining at 1/20000 dilution, shown in red. In Western blot, ab302677 was shown to bind specifically to NR3C1. A band was observed at 94 kDa in wild-type HeLa cell lysates with no signal observed at this size in the NR3C1 knockout cell line. To generate this image, wild-type and NR3C1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a PVDF-FL membrane. Membranes were blocked in Odyssey diluted in equal volume of 0.1 % TBS before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution. Performed under reducing conditions.

All lanes:

Western blot - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] (<a href='/en-us/products/primary-antibodies/glucocorticoid-receptor-antibody-41-glucocorticoid-receptor-ab302677'>ab302677</a>) at 1/1000 dilution

All lanes:

H9 (human lymphoma T lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/10000 dilution

Observed band size: 94 kDa

false

• WB

Supplier Data

Western blot - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] - BSA and Azide Free (AB302678)

This data was developed using ab302677, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. The blot of lane 2 was developed using a high sensitivity ECL substrate. Exposure time : lane 1 : 46 seconds; Lane 2 : 37 seconds.

All lanes:

Western blot - Anti-Glucocorticoid receptor antibody [41/Glucocorticoid Receptor] (<a href='/en-us/products/primary-antibodies/glucocorticoid-receptor-antibody-41-glucocorticoid-receptor-ab302677'>ab302677</a>) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervical adenocarcinoma), whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) (ZB-2305) at 1/10000 dilution

Observed band size: 94 kDa

true