Anti-G3BP antibody [2F3]

• WB

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Western blot - Anti-G3BP antibody [2F3] (AB56574)

G3BP antibody (ab56574) at 1ug/lane + A-431 cell lysate at 25ug/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-G3BP antibody [2F3] (ab56574)

Predicted band size: 52 kDa

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• Flow Cyt

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Flow Cytometry - Anti-G3BP antibody [2F3] (AB56574)

Overlay histogram showing HeLa cells stained with ab56574 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56574, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was generated using the ascites version of the product.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [2F3] (AB56574)

ICC/IF image of ab56574 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab56574, 5μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

This image was generated using the ascites version of the product.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [2F3] (AB56574)

ab56574 staining G3BP in Human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with Triton X-100 and blocked with 5% BSA for 12 hours at 4°C. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 37°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

Top row - untreated cells. Bottom row - cells treated with sodium arsenite. Left - G3BP, Middle - Nucleus, Right - Merge.

Stress granules are visible in cells treated with sodium arsenite, whereas G3BP is dispersed in the cytoplasm in untreated cells.

This image was generated using the ascites version of the product.

This image is courtesy of an anonymous Abreview

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [2F3] (AB56574)

G3BP antibody (ab56574) used in immunohistochemistry at 1ug/ml on formalin fixed and paraffin embedded human lymphoma.

This image was generated using the ascites version of the product.