Anti-RhoA antibody [1B12]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [1B12] (AB54835)

ICC/IF image of ab54835 stained HeLa cells (ab150035). The cells were 4% PFA 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 (ab54835, 1μg/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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA antibody [1B12] (AB54835)

Paraffin embedded human lymphoma tissue stained for RhoA using ab54835 at 5 μg/ml in immunohistochemical analysis.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [1B12] (AB54835)

ab54835 at 10 ug/ml staining RhoA in human Hela cells (ab150035) by Immunocytochemistry / Immunofluorescence.

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

• Flow Cyt

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Flow Cytometry - Anti-RhoA antibody [1B12] (AB54835)

Overlay histogram showing HeLa cells (ab150035) stained with ab54835 (red line). The cells were fixed with methanol (5 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 (ab54835, 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 decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
This image was generated using the ascites version of the product.

• WB

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Western blot - Anti-RhoA antibody [1B12] (AB54835)

RhoA antibody (ab54835) at 1ug/lane + HL-60 cell lysate (ab7914) at 25ug/lane.

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

All lanes:

Western blot - Anti-RhoA antibody [1B12] (ab54835)

Predicted band size: 22 kDa

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• WB

Supplier Data

Western blot - Anti-RhoA antibody [1B12] (AB54835)

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

All lanes:

Western blot - Anti-RhoA antibody [1B12] (ab54835) at 1 µg

Lane 1:

RHOA transfected 293T lysate at 25 µg

Lane 2:

Non-transfected lysate at 25 µg

Secondary

All lanes:

Goat anti-mouse IgG

Predicted band size: 22 kDa

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