Synthetic peptide corresponding to residues in Human IQGAP1.
This product is a recombinant rabbit monoclonal antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.
Our Abpromise guarantee covers the use of ab109292 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/1000 - 1/10000. Detects a band of approximately 195 kDa (predicted molecular weight: 189 kDa).|
|IHC-P||1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Flow Cyt||Use at an assay dependent concentration.|
Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IQGAP1 with unpurified ab109292 at 1/250 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti-rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling IQGAP1 with Purified ab109292 at 1/500 dilution (5 µg/ml). Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: IQGAP1 knockout HAP1 cell lysate (20 µg)
Lane 3: HEK293 cell lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109292 observed at 190 kDa. Red - loading control, ab7291, observed at 52 kDa.
ab109292 was shown to specifically react with IQGAP1 when IQGAP1 knockout samples were used. Wild-type and IQGAP1 knockout samples were subjected to SDS-PAGE. ab109292 and ab7291 (loading control to alpha tubulin) were diluted 1/1000 and 1/2000 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 1/10000 dilution for 1 h at room temperature before imaging.