Anti-G3BP antibody [EPR13986(B)]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling G3BP with purified ab181150 at 1/500 dilution (0.7 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Immunofluorescent analysis of 4% paraformaldehyde fixed 293 cells staining G3BP using ab181150 (unpurified) at 1/100 dilution, and Alexa Fluor®555 stained Goat anti Rabbit IgG at 1/200 dilution as a secondary antibody (red). Dapi counterstain (blue)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling G3BP with purified ab181150 at 1/30 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular cancer tissue sections labeling G3BP with purified ab181150 at 1/50 dilution (6.86 μg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Immunohistochemical analysis of paraffin-embedded Human colon tissue staining G3BP using ab181150 (unpurified) at 1/100 dilution, and prediluted HRP Polymer for Rabbit IgG as a secondary antibody with Hematoxylin counterstain.

• IP

Supplier Data

Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (AB181150)

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

Western blot analysis of G3BP in immunoprecipitation pellets from Ramos cell lysate, using ab181151 (unpurified) at a 1/50 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as a secondary antibody at 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/50 dilution

All lanes:

Immunoprecipitate from Ramos cell lysate using ab181150

Secondary

All lanes:

Peroxidase conjugated Goat anti-Rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 52 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling G3BP with purified ab181150 at 1/50 dilution (6.86 μg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling G3BP with purified ab181150 at 1/50 dilution (6.86 μg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-G3BP antibody [EPR13986(B)] (AB181150)

All lanes:

Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/10000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 20 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 20 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates 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 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 68 kDa

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• WB

Supplier Data

Western blot - Anti-G3BP antibody [EPR13986(B)] (AB181150)

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

All lanes:

Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/5000 dilution

Lane 1:

Jurkat cell lysate at 20 µg

Lane 2:

Ramos cell lysate at 20 µg

Secondary

All lanes:

Peroxidase conjugated Goat anti-Rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 52 kDa

Observed band size: 68 kDa

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• WB

Supplier Data

Western blot - Anti-G3BP antibody [EPR13986(B)] (AB181150)

Lanes 1 - 4 : Merged signal (red and green). Green - ab181150 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab181150 was shown to specifically react with G3BP1 in wild-type A431 cells as signal was lost in G3BP1 knockout cells. Wild-type and G3BP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab181150 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

G3BP1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human G3BP1 knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-g3bp1-knockout-a-431-cell-line-ab261886'>ab261886</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 4:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Predicted band size: 52 kDa

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• IP

Unknown

Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (AB181150)

ab181150 (purified) at 1/20 dilution (2ug) immunoprecipitating G3BP in Ramos whole cell lysate. Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10ug
Lane 2 (+) : ab181150 & Ramos whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab181150 in Ramos whole cell lysate
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150)

Predicted band size: 52 kDa

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