Anti-RhoA antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody (AB86297)

ICC/IF image of ab86297 stained MCF7 cells. 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 (ab86297, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

• WB

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Western blot - Anti-RhoA antibody (AB86297)

All lanes:

Western blot - Anti-RhoA antibody (ab86297) at 1 µg/mL

Lane 1:

Kidney (Mouse) Tissue Lysate at 10 µg

Lane 2:

Western blot - Mouse lung normal tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-lung-normal-tissue-lysate-total-protein-ab29297'>ab29297</a>) at 10 µg

Lane 3:

Kidney (Rat) Tissue Lysate at 10 µg

Lane 4:

Lung (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 22 kDa

Observed band size: 20 kDa

true

Exposure time: 2min

• WB

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Western blot - Anti-RhoA antibody (AB86297)

All lanes:

Western blot - Anti-RhoA antibody (ab86297) at 1 µg/mL

Lane 1:

HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate at 10 µg

Lane 2:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 5:

Human Platelet (Human adult normal cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 22 kDa

Observed band size: 20 kDa

true

Exposure time: 90s

• WB

Unknown

Western blot - Anti-RhoA antibody (AB86297)

All lanes:

Western blot - Anti-RhoA antibody (ab86297) at 1 µg/mL

All lanes:

Recombinant Human RhoA protein (ab101594) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 22 kDa

true

Exposure time: 10s