Anti-CDC42 antibody [EPR15620]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (AB187643)

ab187643 staining CDC42 in Human lung carcinoma tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin-embedded sections). Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on human lung carcinoma.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (AB187643)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling CDC42 with ab187643 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CDC42 antibody [EPR15620] (AB187643)

Intracellular Flow Cytometry analysis of HeLa cells (human cervix adenocarcinoma epithelial).Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Primary antibody was used at a 1/120 dilution (red). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (black). Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody [EPR15620] (AB187643)

Immunocytochemistry/Immunofluorescence analysis of U937 (Human histiocytic lymphoma cell line) labelling CDC42 with purified ab187643 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (AB187643)

ab187643 staining CDC42 in Mouse colon tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed paraffin-embedded sections). Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer pH 9.0). Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on mouse colon.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDC42 antibody [EPR15620] (AB187643)

ab187643 staining CDC42 in Rat colon tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed paraffin-embedded sections). Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer pH 9.0). Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on rat colon.

• WB

Lab

Western blot - Anti-CDC42 antibody [EPR15620] (AB187643)

Lanes 1-4 : Merged signal (red and green). Green - ab187643 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.

ab187643 Anti-CDC42 antibody [EPR15620] was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab187643 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution

Lane 1:

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

Lane 2:

CDC42 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human CDC42 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-cdc42-knockout-hek-293t-cell-line-ab266522'>ab266522</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 21 kDa

Observed band size: 20 kDa

false

• WB

Lab

Western blot - Anti-CDC42 antibody [EPR15620] (AB187643)

Lanes 1 - 4 : Merged signal (red and green). Green - ab187643 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab187643 was shown to specifically react with CDC42 in wild-type HAP1 cells. No band was observed when CDC42 knockout samples were examined. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab187643 and ab8245 (loading control to GAPDH) were diluted at 1/5000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

CDC42 knockout HAP1 cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-CDC42 antibody [EPR15620] (AB187643)

All lanes:

Western blot - Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat spleen lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Supplier Data

Western blot - Anti-CDC42 antibody [EPR15620] (AB187643)

All lanes:

Western blot - Anti-CDC42 antibody [EPR15620] (ab187643) at 1/20000 dilution

Lane 1:

Jurkat cell lysate at 20 µg

Lane 2:

HT-29 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false