Anti-GNAS antibody [EPR24177-24]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAS antibody [EPR24177-24] (AB283266)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling GNAS with ab283266 at 1/1000 (0.473 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat cerebrum. The section was incubated with ab283266 for 10 mins at room temperature. 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) was used.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAS antibody [EPR24177-24] (AB283266)

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labelling GNAS with ab283266 at 1/2500 (0.189 μg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human ovarian cancer. The section was incubated with ab283266 for 10 mins at room temperature. 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) was used.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GNAS antibody [EPR24177-24] (AB283266)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 cells labelling GNAS with ab283266 at 1/2000 (0.237 μg/mL) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 μg/mL) dilution (Green). Confocal image showing membranous staining in MCF7 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 μg/mL) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/mL) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAS antibody [EPR24177-24] (AB283266)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling GNAS with ab283266 at 1/1000 (0.473 μg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebrum. The section was incubated with ab283266 for 10 mins at room temperature. 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) was used.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAS antibody [EPR24177-24] (AB283266)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling GNAS with ab283266 at 1/2500 (0.189 μg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human normal cerebrum. The section was incubated with ab283266 for 10 mins at room temperature.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) was used.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-GNAS antibody [EPR24177-24] (AB283266)

GNAS was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab283266 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283266 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg.

Lane 2 : ab283266 IP in MCF7 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283266 in MCF7 whole cell lysate.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-GNAS antibody [EPR24177-24] (ab283266)

Predicted band size: 46 kDa

Observed band size: 44 kDa

false

• WB

Lab

Western blot - Anti-GNAS antibody [EPR24177-24] (AB283266)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The near two bands represent two different isoforms. The expression profile/molecular weight observed are consistent with what has been described in the literature (PMID : 3258290).

Exposure time : 15 seconds.

All lanes:

Western blot - Anti-GNAS antibody [EPR24177-24] (ab283266) at 1/1000 dilution

All lanes:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 46 kDa

Observed band size: 44 kDa

false

• WB

Lab

Western blot - Anti-GNAS antibody [EPR24177-24] (AB283266)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTThe near two bands represent two different isoforms. The expression profile/molecular weight observed are consistent with what has been described in the literature (PMID : 3258290).

Exposure time : Lane 1, 3 : 37 seconds Lane 2 : 59 seconds.

All lanes:

Western blot - Anti-GNAS antibody [EPR24177-24] (ab283266) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 44 kDa

false

• WB

Lab

Western blot - Anti-GNAS antibody [EPR24177-24] (AB283266)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The near two bands represent two different isoforms. The expression profile/molecular weight observed are consistent with what has been described in the literature (PMID : 3258290).

Exposure time : Lane 1-7 : 37 seconds Lane 8 : 15 seconds.

All lanes:

Western blot - Anti-GNAS antibody [EPR24177-24] (ab283266) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate 20 at 20 µg

Lane 4:

MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat hippocampus tissue lysate at 20 µg

Lane 7:

Mouse brain tissue lysate at 20 µg

Lane 8:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 8:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lane 8:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 44 kDa

false