WB: Human placenta tissue lysate and HeLa and HEK293 whole cell lysates.
IHC-P: Human skin melanoma tissue.
ICC/IF: MCF7 cells.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 205 kDa (predicted molecular weight: 189 kDa).
Use a concentration of 1 µg/ml.
Binds to activated CDC42 but does not stimulate its GTPase activity. It associates with calmodulin. Could serve as an assembly scaffold for the organization of a multimolecular complex that would interface incoming signals to the reorganization of the actin cytoskeleton at the plasma membrane. May promote neurite outgrowth.
Expressed in the placenta, lung, and kidney. A lower level expression is seen in the heart, liver, skeletal muscle and pancreas.
Regions C1 and C2 can either interact with nucleotide-free CDC42, or interact together, depending on the phosphorylation state of Ser-1443. When Ser-1443 is not phosphorylated, C1 and C2 interact, which prevents binding of nucleotide-free CDC42 and promotes binding of GTP-bound CDC42. Phosphorylation of Ser-1443 prevents interaction between C1 and C2, which opens the structure of the C-terminus and allows binding and sequestration of nucleotide-free CDC42 on both C1 and C2.
Phosphorylation of Ser-1443 by PKC prevents interaction between C1 and C2, allowing binding of nucleotide-free CDC42. Ser-1443 phosphorylation enhances the ability to promote neurite outgrowth.
IQ motif containing GTPase activating protein 1 antibody
Ras GTPase activating like protein 1 antibody
Ras GTPase-activating-like protein IQGAP1 antibody
RasGAP-like with IQ motifs antibody
Western blot - Anti-IQGAP1 antibody (ab110203)
Predicted band size: 189 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: IQGAP1 knockout HAP1 cell lysate (20 µg) Lane 3: HeLa cell lysate (20 µg) Lane 4: HEK293 cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab110203 observed at 190 kDa. Red - loading control, ab7291, observed at 52 kDa. ab110203 was shown to specifically react with IQGAP1 when IQGAP1 knockout samples were used. Wild-type and IQGAP1 knockout samples were subjected to SDS-PAGE. ab110203 and ab7291 (loading control to alpha tubulin) were diluted 1 μg/mL and 1/2000 respectively and incubated overnight at 4°C. Blots were developed withGoat 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.
ICC/IF image of ab110203 stained MCF7 cells. The cells were 4% formaldehyde 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 (ab110203 1µg/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. This antibody also gave a positive result in 4% foarmaldehyde fixed (10 min) Hek293 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa and MCF7 cells at 1µg/ml.
Western blot - Anti-IQGAP1 antibody (ab110203)
All lanes : Anti-IQGAP1 antibody (ab110203) at 1 µg/ml
Lane 1 : Human placenta tissue lysate - total protein (ab29745) Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
IHC image of IQGAP1 staining in Human skin melanoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110203, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.