Anti-Glucocorticoid Receptor antibody [2153CT307.32.80] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Glucocorticoid Receptor antibody [2153CT307.32.80] - BSA and Azide free (AB307606)

This data was developed using ab307605, the same antibody clone in a different buffer formulation.

A549 wild-type and Glucocorticoid Receptor knockout cells were labelled with a green or violet, fluorescent dye, respectively. Wild-type and knockout cells were mixed in a 1 : 1 ratio, fixed in 4% PFA and permeabilized in 0.1% Triton X-100.

Antibody staining was quantified using the Attune NxT Flow Cytometer with representative images showing the staining intensity in the knockout population (pink histogram, dashed line) compared to the wild-type cells (green histogram, solid line). Histograms with dotted lines represent secondary antibody-only controls in both wild-type and knockout cells.

Antibody dilution : 1/10260

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.

• WB

Supplier Data

Western blot - Anti-Glucocorticoid Receptor antibody [2153CT307.32.80] - BSA and Azide free (AB307606)

This data was developed using ab307605, the same antibody clone in a different buffer formulation.

ab307605 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 ab307605 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 [2153CT307.32.80] (<a href='/ko/products/primary-antibodies/glucocorticoid-receptor-antibody-2153ct3073280-ab307605'>ab307605</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

false

• WB

Supplier Data

Western blot - Anti-Glucocorticoid Receptor antibody [2153CT307.32.80] - BSA and Azide free (AB307606)

This data was produced using ab307605, the same antibody clone but in a different buffer.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Glucocorticoid Receptor antibody [2153CT307.32.80] (<a href='/ko/products/primary-antibodies/glucocorticoid-receptor-antibody-2153ct3073280-ab307605'>ab307605</a>) at 1/1000 dilution

All lanes:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 85 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Glucocorticoid Receptor antibody [2153CT307.32.80] - BSA and Azide free (AB307606)

This data was produced using ab307605, the same antibody clone but in a different buffer.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab307605 was shown to bind specifically to Glucocorticoid Receptor. A band was observed at 85 kDa in wild-type HeLa cell lysates with whereas no signal observed at this size in Glucocorticoid Receptor knockout cell line ab261766 (knockout cell lysate ab257009).

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Glucocorticoid Receptor antibody [2153CT307.32.80] (<a href='/ko/products/primary-antibodies/glucocorticoid-receptor-antibody-2153ct3073280-ab307605'>ab307605</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NR3C1 (Glucocorticoid Receptor) knockout HeLa cell lysate (<a href='/ko/products/cell-lysates/human-nr3c1-glucocorticoid-receptor-knockout-hela-cell-lysate-ab257009'>ab257009</a>) at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Observed band size: 85 kDa

true

Exposure time: 136s