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    products/primary-antibodies/iqgap1-antibody-epr5221-ab109292.pdf

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Signal Transduction Adapters Cytoplasmic
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IQGAP1 antibody [EPR5221] (ab109292)

  • Datasheet
  • SDS
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Western blot - Anti-IQGAP1 antibody [EPR5221] (ab109292)
  • Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5221] (ab109292)
  • Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody [EPR5221] (ab109292)
  • Western blot - Anti-IQGAP1 antibody [EPR5221] (ab109292)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody [EPR5221] (ab109292)
  • Anti-IQGAP1 antibody [EPR5221] (ab109292)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5221] to IQGAP1
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human, African green monkey

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Carrier Free

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Overview

  • Product name

    Anti-IQGAP1 antibody [EPR5221]
    See all IQGAP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR5221] to IQGAP1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cyt (Intra)more details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human, African green monkey
  • Immunogen

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

  • Positive control

    • HeLa, COS-1, HUVEC, and Jurkat cell lysates, Human uterus 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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5221
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • Small G Proteins
    • Regulators
    • Cancer
    • Signal transduction
    • Other

Associated products

  • Alternative Versions

    • Anti-IQGAP1 antibody [EPR5221] - BSA and Azide free (ab247821)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa membrane extract lysate (ab29547)
    • Jurkat membrane extract lysate (ab30130)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
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 (Intra)
Use at an assay dependent concentration.
Notes
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 (Intra)
Use at an assay dependent concentration.
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.
  • Target information above from: UniProt accession P46940 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8826 Human
    • Omim: 603379 Human
    • SwissProt: P46940 Human
    • Unigene: 430551 Human
    • 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 [EPR5221] (ab109292)
      Western blot - Anti-IQGAP1 antibody [EPR5221] (ab109292)

      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.

    • Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5221] (ab109292)
      Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5221] (ab109292)

      Intracellular 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 - Anti-IQGAP1 antibody [EPR5221] (ab109292)
      Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody [EPR5221] (ab109292)

      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.

    • Western blot - Anti-IQGAP1 antibody [EPR5221] (ab109292)
      Western blot - Anti-IQGAP1 antibody [EPR5221] (ab109292)
      All lanes : Anti-IQGAP1 antibody [EPR5221] (ab109292) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : Cos-1 cell lysate
      Lane 3 : HUVEC cell lysate
      Lane 4 : Jurkat cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 189 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody [EPR5221] (ab109292)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody [EPR5221] (ab109292)

      Immunohistochemical analysis of paraffin-embedded Human uterus tissue using ab109292 at a dilution of 1/500.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Anti-IQGAP1 antibody [EPR5221] (ab109292)
      Anti-IQGAP1 antibody [EPR5221] (ab109292)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (4)

    Publishing research using ab109292? Please let us know so that we can cite the reference in this datasheet.

    ab109292 has been referenced in 4 publications.

    • Chen A  et al. Inhibition of polar actin assembly by astral microtubules is required for cytokinesis. Nat Commun 12:2409 (2021). PubMed: 33893302
    • Chen A  et al. The scaffold-protein IQGAP1 enhances and spatially restricts the actin-nucleating activity of Diaphanous-related formin 1 (DIAPH1). J Biol Chem 295:3134-3147 (2020). PubMed: 32005666
    • Katsumura KR  et al. Human leukemia mutations corrupt but do not abrogate GATA-2 function. Proc Natl Acad Sci U S A 115:E10109-E10118 (2018). PubMed: 30301799
    • Tekletsadik YK  et al. A conserved role of IQGAP1 in regulating TOR complex 1. J Cell Sci 125:2041-52 (2012). ICC/IF ; Human . PubMed: 22328503

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