Anti-RhoGDI antibody [2G3]

• WB

Lab

Western blot - Anti-RhoGDI antibody [2G3] (AB135252)

Western blot : Anti-ARHGDIA antibody [2G3] (ab135252) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab135252 was shown to bind specifically to ARHGDIA. A band was observed at 27 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in ARHGDIA knockout cell line ab266446 (knockout cell lysate ab257355). To generate this image, wild-type and ARHGDIA knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-RhoGDI antibody [2G3] (ab135252) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

ARHGDIA knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-arhgdia-rhogdi-knockout-hek-293t-cell-line-ab266446'>ab266446</a>)

Lane 2:

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-arhgdia-rhogdi-knockout-hek-293t-cell-lysate-ab257355'>ab257355</a>)

Lane 3:

Wild-type HEK-293T ab255553 cell lysate at 20 µg

Lane 4:

ARHGDIA knockout HEK-293T ab263544 cell lysate at 20 µg

Lane 5:

HeLa cell lysate at 20 µg

Lane 6:

U-87 MG cell lysate at 20 µg

Lane 7:

Human Stomach cell lysate at 20 µg

Lane 8:

Human Skeletal Muscle cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 27 kDa

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• Flow Cyt

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Flow Cytometry - Anti-RhoGDI antibody [2G3] (AB135252)

Flow cytometric analysis of HeLa cells using ab135252 at 1/200 dilution (green) and negative control (red).

• ICC/IF

AbReview32693****

Immunocytochemistry/ Immunofluorescence - Anti-RhoGDI antibody [2G3] (AB135252)

Immunofluorescence analysis of rat lung epithelial cells, staining RhoGDI with ab135252.

Cells were fixed with paraformaldehyde, permeabilized with 0.2% saponin and blocked with 10% serum for 10 minutes at 20°C. Samples were incubated with primary antibody (1/200 in 10% serum, 0.2% saponin in PBS) for 1 hour at 23°C. An AlexaFluor®488-conjugated goat anti-mouse polyclonal IgG (1/400) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RhoGDI antibody [2G3] (AB135252)

Immunofluorescence analysis of HepG2 cells using ab135252 at 1/200 dilution (green). Blue : DRAQ5 fluorescent DNA dye.

• WB

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Western blot - Anti-RhoGDI antibody [2G3] (AB135252)

All lanes:

Western blot - Anti-RhoGDI antibody [2G3] (ab135252) at 1/500 dilution

Lane 1:

Jurkat cell lysate

Lane 2:

HeLa cell lysate

Lane 3:

NIH3T3 cell lysate

Lane 4:

C6 (Rat glial tumor cell line) cell lysate

Lane 5:

K562 cell lysate

Lane 6:

COS-7 (African green monkey kidney fibroblast-like cell line) cell lysate

Predicted band size: 23 kDa

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• WB

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Western blot - Anti-RhoGDI antibody [2G3] (AB135252)

Note : the predicted band size of the full length protein is 23 kDa. The expected band size of the recombinant protein used here is 42.7 kDa.

All lanes:

Western blot - Anti-RhoGDI antibody [2G3] (ab135252) at 1/500 dilution

All lanes:

Human RhoGDI recombinant protein.

Predicted band size: 23 kDa

Observed band size: 48.7 kDa

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