Recombinant Anti-CDC42 antibody [EPR15620] - BSA and Azide free (ab271953)

Lane 1 (input): HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate,10μg
Lane 2(+): HT-29 whole cell lysate
Lane 3(-): Rabbit monoclonal IgG (ab172730) instead of  ab187643 in HT-29 whole cell lysate

Ab187643 immunoprecipitating CDC42 in HT-29 whole cell lysate. Capture antibody was used at a 1:60 dilution. For western blotting, ab187643 was used as the primary antibody at a 1:1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Blocking and diluting buffer: 5% NFDM/TBST

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). 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

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.

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

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.