Anti-G3BP antibody [EPR13985(B)]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13985(B)] (AB181149)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling G3BP with Purified ab181149 at 1 : 300 (0.4 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13985(B)] (AB181149)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling G3BP with Purified ab181149 at 1 : 300 (0.4 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• Flow Cyt

Lab

Flow Cytometry - Anti-G3BP antibody [EPR13985(B)] (AB181149)

Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling G3BP with Purified ab181149 at 1 : 20 dilution (5 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13985(B)] (AB181149)

Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling G3BP with Purified ab181149 at 1 : 50 dilution (2.4 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]+H21 : L21 - Microtubule Marker (Alexa Fluor® 594) 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.

• IP

Lab

Immunoprecipitation - Anti-G3BP antibody [EPR13985(B)] (AB181149)

Purified ab181149 at 1 : 20 dilution (0.6μg) immunoprecipitating G3BP in Ramos whole cell lysate.
Lane 1 (input) : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 μg.
Lane 2 (+) : ab181149 + Ramos whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab181149 in Ramos whole cell lysate.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1 : 1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : kDa

All lanes:

Immunoprecipitation - Anti-G3BP antibody [EPR13985(B)] (ab181149)

Predicted band size: 52 kDa

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• WB

Supplier Data

Western blot - Anti-G3BP antibody [EPR13985(B)] (AB181149)

All lanes:

Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149) at 1/20000 dilution

Lane 1:

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

Lane 2:

Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 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

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• WB

Lab

Western blot - Anti-G3BP antibody [EPR13985(B)] (AB181149)

Lanes 1 - 4 : Merged signal (red and green). Green - ab181149 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab181149 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. ab181149 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 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 [EPR13985(B)] (ab181149) at 1/10000 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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