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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

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Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
  • Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
  • Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
  • Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
  • Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
  • Flow Cytometry (Intracellular) - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
  • Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17963] to GEF H1 - C-terminal
  • Suitable for: ICC/IF, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-GEF H1 antibody [EPR17963] - C-terminal
    See all GEF H1 primary antibodies

  • Description

    Rabbit monoclonal [EPR17963] to GEF H1 - C-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, HEK293, C6 and NIH/3T3 cell lysate. ICC/IF: HeLa and MCF-7 cells. Flow Cyt (intra): HEK293 cells.

  • 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab201687 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
1/500.
WB
1/2000. Detects a band of approximately 112 kDa (predicted molecular weight: 112 kDa).
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Notes
ICC/IF
1/500.
WB
1/2000. Detects a band of approximately 112 kDa (predicted molecular weight: 112 kDa).
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Activates Rho-GTPases by promoting the exchange of GDP for GTP. May be involved in epithelial barrier permeability, cell motility and polarization, dendritic spine morphology, antigen presentation, leukemic cell differentiation, cell cycle regulation, and cancer. Binds Rac-GTPases, but does not seem to promote nucleotide exchange activity toward Rac-GTPases, which was uniquely reported in PubMed:9857026. May stimulate instead the cortical activity of Rac. Inactive toward CDC42, TC10, or Ras-GTPases. Forms an intracellular sensing system along with NOD1 for the detection of microbial effectors during cell invasion by pathogens. Required for RHOA and RIP2 dependent NF-kappaB signaling pathways activation upon S.flexneri cell invasion. Involved not only in sensing peptidoglycan (PGN)-derived muropeptides through NOD1 that is independent of its GEF activity, but also in the activation of NF-kappaB by Shigella effector proteins (IpgB2 and OspB) which requires its GEF activity and the activation of RhoA.

  • Sequence similarities

    Contains 1 DH (DBL-homology) domain.
    Contains 1 PH domain.
    Contains 1 phorbol-ester/DAG-type zinc finger.

  • Domain

    The DH (DBL-homology) domain interacts with and promotes loading of GTP on RhoA.
    The PH (pleckstrin-homology) domain is involved in microtubule binding and targeting to tight junctions.

  • Post-translational
    modifications

    Phosphorylation of Ser-886 by PAK1 induces binding to protein 14-3-3 zeta, promoting its relocation to microtubules and the inhibition of its activity. Phosphorylated by STK6 and CDK1 during mitosis, which negatively regulates its activity. Phosphorylation by MAPK1 or MAPK3 increases nucleotide exchange activity. Phosphorylation by PAK4 releases GEF-H1 from the microtubules.

  • Cellular localization

    Cytoplasm. Cell junction > tight junction. Golgi apparatus. Cytoplasm > cytoskeleton > spindle. Cell projection > ruffle membrane. Localizes to the tips of cortical microtubules of the mitotic spindle during cell division, and is further released upon microtubule depolymerization. Recruited into membrane ruffles induced by S.flexneri at tight junctions of polarized epithelial cells.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • AA408978 antibody
    • ARHG2 antibody
    • ARHG2_HUMAN antibody
    • ARHGEF 2 antibody
    • ARHGEF-2 antibody
    • ARHGEF2 antibody
    • GEF antibody
    • GEF H1 antibody
    • GEF-H1 antibody
    • GEFH1 antibody
    • Guanine nucleotide exchange factor H1 antibody
    • KIAA0651 antibody
    • Lbcl1 antibody
    • Lfc antibody
    • LFP40 antibody
    • MGC95068 antibody
    • Microtubule-regulated Rho-GEF antibody
    • mKIAA0651 antibody
    • P40 antibody
    • Proliferating cell nucleolar antigen p40 antibody
    • Protein GEF-H1 antibody
    • Rho guanine nucleotide exchange factor 2 antibody
    • rho/rac guanine nucleotide exchange factor (GEF) 2 antibody
    • rho/rac guanine nucleotide exchange factor 2 antibody
    • rho/rac guanine nucleotide exchange factor antibody
    see all

Images

  • Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    All lanes : Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ARHGEF knockout HeLa cell lysate
    Lane 3 : HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 112 kDa
    Observed band size: 112 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab201687 observed at 112 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab201687 Anti-GEF H1 antibody [EPR17963] - C-terminal was shown to specifically react with GEF H1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265797 (knockout cell lysate ab257223) was used. Wild-type and GEF H1 knockout samples were subjected to SDS-PAGE. ab201687 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 10000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    All lanes : Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/10000 dilution

    Lane 1 : HEK293 (Human embryonic kidney) lysate
    Lane 2 : HEK293 (Human epithelial cells from cervix adenocarcinoma) lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 112 kDa
    Observed band size: 112 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    All lanes : Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate
    Lane 3 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) lysate
    Lane 4 : C6 (Rat glial tumor cells) lysate
    Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 112 kDa
    Observed band size: 112 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling GEF H1 with ab201687 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). 

    Confocal image showing cytoplasmic staining on HeLa cell line is observed.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab201687 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling GEF H1 with ab201687 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic staining on MCF7 cell line is observed.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab201687 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Flow Cytometry (Intracellular) - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    Flow Cytometry (Intracellular) - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

    Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HEK293 (human embryonic kidney) cells labeling GEF H1 with ab201687 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody. 

     

     

  • Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)
    Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

References (2)

Publishing research using ab201687? Please let us know so that we can cite the reference in this datasheet.

ab201687 has been referenced in 2 publications.

  • He L  et al. Actin-granule formation is an additional step in cardiac myofibroblast differentiation. Ann Transl Med 9:165 (2021). PubMed: 33569467
  • Chan DCH  et al. Arhgef2 regulates mitotic spindle orientation in hematopoietic stem cells and is essential for productive hematopoiesis. Blood Adv 5:3120-3133 (2021). PubMed: 34406376

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