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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IQGAP1 antibody [EPR5220] (ab133490)

Submit a review Submit a question References (10)
Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490)
  • Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody [EPR5220] (ab133490)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody [EPR5220] (ab133490)
  • Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5220] (ab133490)
  • Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490)
  • Anti-IQGAP1 antibody [EPR5220] (ab133490)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5220] to IQGAP1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 APC Carrier Free HRP PE

Overview

  • Product name

    Anti-IQGAP1 antibody [EPR5220]
    See all IQGAP1 primary antibodies

  • Description

    Rabbit monoclonal [EPR5220] to IQGAP1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • HeLa, 293T, Human placenta, Mouse thymus and F9 cell lysates; Human kidney tissue

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133490 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  
WB
1/1000 - 1/10000. Detects a band of approximately 195 kDa (predicted molecular weight: 189 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50 - 1/100.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  
WB
1/1000 - 1/10000. Detects a band of approximately 195 kDa (predicted molecular weight: 189 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50 - 1/100.
Application notes
Is unsuitable for IP.

Target

  • Function

    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.

  • Tissue specificity

    Expressed in the placenta, lung, and kidney. A lower level expression is seen in the heart, liver, skeletal muscle and pancreas.

  • Sequence similarities

    Contains 1 CH (calponin-homology) domain.
    Contains 4 IQ domains.
    Contains 1 Ras-GAP domain.
    Contains 1 WW domain.

  • Domain

    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.

  • Post-translational
    modifications

    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.

  • Cellular localization

    Cell membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • HUMORFA01 antibody
      • IQ motif containing GTPase activating protein 1 antibody
      • IQGA1_HUMAN antibody
      • Iqgap1 antibody
      • KIAA0051 antibody
      • p195 antibody
      • Ras GTPase activating like protein 1 antibody
      • Ras GTPase-activating-like protein IQGAP1 antibody
      • RasGAP-like with IQ motifs antibody
      • SAR1 antibody
      see all

    Images

    • Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490)
      Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: IQGAP 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 - ab133490 observed at 190 kDa. Red - loading control, ab7291, observed at 52 kDa.
      ab133490 was shown to specifically react with IQGAP1 when IQGAP1 knockout samples were used. Wild-type and IQGAP1 knockout samples were subjected to SDS-PAGE. ab133490 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 1/10 000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody [EPR5220] (ab133490)
      Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody [EPR5220] (ab133490)

      Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling IQGAP1 with Purified ab133490 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody [EPR5220] (ab133490)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody [EPR5220] (ab133490)

      Immunohistochemical analysis of IQGAP1 in paraffin embedded Human kidney tissue, using ab133490 at a dilution of 1/100.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5220] (ab133490)
      Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5220] (ab133490)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IQGAP1 with unpurified ab133490 at 1/20 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. 

       

       

    • Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490)
      Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490)
      All lanes : Anti-IQGAP1 antibody [EPR5220] (ab133490) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : Human placenta lysate
      Lane 4 : Mouse thymus lysate
      Lane 5 : F9 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 189 kDa
      Observed band size: 195 kDa why is the actual band size different from the predicted?

    • Anti-IQGAP1 antibody [EPR5220] (ab133490)
      Anti-IQGAP1 antibody [EPR5220] (ab133490)

    References (10)

    Publishing research using ab133490? Please let us know so that we can cite the reference in this datasheet.

    ab133490 has been referenced in 10 publications.

    • Hebert JD  et al. The scaffold protein IQGAP1 is crucial for extravasation and metastasis. Sci Rep 10:2439 (2020). PubMed: 32051509
    • Sabo Y  et al. IQGAP1 Negatively Regulates HIV-1 Gag Trafficking and Virion Production. Cell Rep 30:4065-4081.e4 (2020). PubMed: 32209469
    • He C  et al. Engineering of HN3 increases the tumor targeting specificity of exosomes and upgrade the anti-tumor effect of sorafenib on HuH-7 cells. PeerJ 8:e9524 (2020). PubMed: 33062407
    • Li S  et al. Follicle-stimulating hormone promoted pyruvate kinase isozyme type M2-induced glycolysis and proliferation of ovarian cancer cells. Arch Gynecol Obstet 299:1443-1451 (2019). PubMed: 30809696
    • Fang Y  et al. The adhesion and migration of microglia to ß-amyloid (Aß) is decreased with aging and inhibited by Nogo/NgR pathway. J Neuroinflammation 15:210 (2018). PubMed: 30029608
    • Jin X  et al. Dysregulation of INF2-mediated mitochondrial fission in SPOP-mutated prostate cancer. PLoS Genet 13:e1006748 (2017). Human . PubMed: 28448495
    • Diao B  et al. IQGAP1-siRNA inhibits proliferation and metastasis of U251 and U373 glioma cell lines. Mol Med Rep 15:2074-2082 (2017). WB . PubMed: 28259970
    • Cui X  et al. Elevated IQGAP1 and CDC42 levels correlate with tumor malignancy of human glioma. Oncol Rep 37:768-776 (2017). WB, IHC . PubMed: 28035419
    • Keshari PK  et al. Allelic imbalance of multiple sclerosis susceptibility genes IKZF3 and IQGAP1 in human peripheral blood. BMC Genet 17:59 (2016). WB . PubMed: 27080863
    • Jin X  et al. The Overexpression of IQGAP1 and ß-Catenin Is Associated with Tumor Progression in Hepatocellular Carcinoma In Vitro and In Vivo. PLoS One 10:e0133770 (2015). IHC, WB ; Human . PubMed: 26252773

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