Overview

  • Product name

    Anti-GEF H1 antibody [B4/7]
    See all GEF H1 primary antibodies
  • Description

    Mouse monoclonal [B4/7] to GEF H1
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-P, WB, IP, IHC-Fr, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Dog, Human
  • Immunogen

    Protein fraction isolated from detergent extracts of MDCK cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab90783 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
IHC-P Use a concentration of 5 µg/ml.
WB 1/50. Predicted molecular weight: 112 kDa.
IP Use at an assay dependent concentration.
IHC-Fr 1/50.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, 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

  • 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

  • IHC image of ab90783 staining in Hu breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab90783, 5µg/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.
  • Overlay histogram showing HeLa cells stained with ab90783 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab90783, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:

  • Cao J  et al. Increased expression of GEF-H1 promotes colon cancer progression by RhoA signaling. Pathol Res Pract 215:1012-1019 (2019). Read more (PubMed: 30846413) »
  • Yu J  et al. Wnt5a induces ROR1/ROR2 heterooligomerization to enhance leukemia chemotaxis and proliferation. J Clin Invest 126:585-98 (2016). Read more (PubMed: 26690702) »
See all 3 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (brain)
Specification
brain
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: microwave
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: rt°C

Abcam user community

Verified customer

Submitted Feb 15 2012

Answer

Thank you for your phone call.

I am sorry to hear that ab90783 did not work in rat in IHC with PFA fixed sections. Since this species and IHC technique had not been tested, I'd be happy to offer you a testing discount retrospectively as a one-time compensation.


DISCOUNT CODE: xxx for ab90783
DISCOUNT CODE: xxx for ab94348
Expiration date: xxx

Also, I am very pleased to hear you would like to accept our offer and test ab94348 in rat and IHC (PFA fixed).

Each code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for rat and/or IHC (PFA fixed) and include this code in the “Additional Comments” section so we know the Abreview is for this promotion.

For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. Each code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your reply and your patience. I have contacted the laboratory producing the ab103558 in regards to the recognition of the phospho and the non-phospho form. The lab replied that there is a residue which can be reportedly phosphorylated within the epitope. Unfortunately we have no data upon which to judge the relative efficiency of binding of the antibody to the phosphorylated vs non-phosphorylated epitope. However as the immunogen is different from the immunogen of the ab90783, there might not be the same pattern? The immunogen for ab103558 is region between residue 625-675. We do not have small aliquots which we could provide for testing purposes. However I can make you a testing offer, if you would need to test this antibody also on another application or species? For example on frozen section immunohistochemistry? The offer would consist of a discount of a future antibody purchase after you would have provided the feedback. The discount would be 100%. Please do let me know if you would be interested, as I would need to provide you a code. I am sorry for not being more helpful at the moment. Please let me know if I could help you otherwise or if you would like to discuss the results of ab90783. I am looking forward to hear back from you and wish you meanwhile also a very merry Christmas time!

Read More

Answer

Thank you for contacting us. I am sorry to hear that you are experiencing problems with ab90783. To my knowledge, the epitope of the ab90783 has not been determined. It is a monoclonal antibody generated against a cell fraction. I have not been aware that the ab90783 does not recognize the phosphorylated form. Can you please explain in more details what results have been obtained with the ab90783 compared to the ab94348? Do you have any staining at all with the ab90783? As antibodies are biological material, it can happen that they are damaged during transport or storage for example. I would appreciate if you could provide more details, including eventually an image, and I could investigate whether we can also replace this antibody vial for you. In regard to the epitopes of the other antibodies: -ab94348 has been generated against a phosphorylated peptide around S885 -ab79175 has also been generated against a non-phosphopeptide around S885, Detects endogenous levels of total GEF H1 - I do not know the exact sequence, as this is proprietary information for this antibody. -ab103558 has been generated against a peptide corresponding to a region between amino acid 625-675. I do not know the exact sequence, as this is proprietary information for this antibody. I hope this information is helpful to you. I am looking forward to hear back from you.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (293 cells)
Loading amount
30 µg
Specification
293 cells
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Melanie Blasius

Verified customer

Submitted Nov 26 2010

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293 cells)
Total protein in input
1000 µg
Specification
293 cells
Immuno-precipitation step
Other - protein G dynabeads

Dr. Melanie Blasius

Verified customer

Submitted Nov 26 2010

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