Overview

  • Product nameAnti-RhoGAP antibody [EP489Y]
    See all RhoGAP primary antibodies
  • Description
    Rabbit monoclonal [EP489Y] to RhoGAP
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (Human) from RhoGAP.

  • Positive control
    • Human stomach carcinoma and the following cell lysates: Hela, NIH 3T3, Jurkat, 293, K562, MCF-7, A431 and PC12.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • PurityProtein A purified
  • ClonalityMonoclonal
  • Clone numberEP489Y
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32328 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
WB 1/500. Detects a band of approximately 190 kDa (predicted molecular weight: 171 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/60 - 1/1000. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
  • Application notesIs unsuitable for ICC or IP.
  • Target

    • RelevanceThe p190 RhoGAP protein is associated with p120 RasGAP in growth-factor stimulated and tyrosine kinase transformed cells. It functions as a GTPase - activating protein (GAP) for Rho and Rac family proteins, which are involved in regulating cytoskeletal actin and membrane ruffling. The antibody appears to couple signal transduction via Ras and Rho through its association with RasGAP. The presence of two adjacent SH2 domains in the p21 RAS GAP indicates that GAP might interact directly with tyrosine kinases. p190B can be directly phosphorylated on a single identified tyrosine residue by the activated insulin and IGF-1 receptors. This phosphorylation causes p190B to translocate to the lipid rafts of the plasma membrane, where it can facilitate the inactivation of the Rho GTPase. P190B also has been shown to be tyrosine phosphorylated by cSrc and vSrc. Additionally the GAP domain of p190B is shown to attenuate signal transducing activity of Rac, Rho and CDC42.
    • Cellular localizationCytoplasmic
    • Database links
    • Alternative names
      • ARHGAP 5 antibody
      • ARHGAP5 antibody
      • p190 B antibody
      • p190 RhoGAP antibody
      • Rho GTPase activating protein 5 antibody
      • RhoGAP 5 antibody
      • RhoGAP antibody
      • RhoGAP5 antibody
      see all

    Anti-RhoGAP antibody [EP489Y] images

    • Overlay histogram showing A431 cells stained with ab32328 (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 (ab32328, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Anti-RhoGAP antibody [EP489Y] (ab32328) at 1/500 dilution + Mouse MEFs whole cell lysate at 50 µg

      Secondary
      HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 171 kDa
      Observed band size : 190 kDa (why is the actual band size different from the predicted?)


      Exposure time : 5 minutes

      This image is courtesy of an anonymous Abreview

      See Abreview

    • All lanes : Anti-RhoGAP antibody [EP489Y] (ab32328) at 1/500 dilution

      Lane 1 : Hela cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : MCF-7 cell lysate
      Lane 4 : K562 cell lysate
      Lane 5 : A431 cell lysate
      Lane 6 : 293 cell lysate
      Lane 7 : NIH 3T3 cell lysate
      Lane 8 : PC12 cell lysate


      Predicted band size : 171 kDa
      Observed band size : 190 kDa (why is the actual band size different from the predicted?)
    • ab32328, at a 1/100 dilution, staining human RhoGAP in stomach carcinoma by Immunohistochemistry, Paraffin embedded tissue

    References for Anti-RhoGAP antibody [EP489Y] (ab32328)

    This product has been referenced in:
    • Repetto D  et al. p140Cap regulates memory and synaptic plasticity through Src-mediated and citron-N-mediated actin reorganization. J Neurosci 34:1542-53 (2014). Mouse . Read more (PubMed: 24453341) »

    See 1 Publication for this product

    Product Wall

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
    Sample Human Tissue sections (Human placenta)
    Specification Human placenta
    Permeabilization Yes - Triton 1%
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Dec 03 2014

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 10µg/mL · Temperature: RT°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
    Sample Dog Tissue sections (MDCK pellets in paraffin)
    Specification MDCK pellets in paraffin
    Permeabilization No
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Oct 13 2014

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Sample Human Cell (293T)
    Specification 293T
    Permeabilization Yes - Tween 20, 0,1%
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted May 13 2014

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (10)
    Sample Mouse Cell lysate - whole cell (MEFs)
    Specification MEFs
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Dec 16 2013

    Thank you for you enquiry regarding ab32328. This antibody generated against sequence from p190 RhoGAP B isoform and we have not tested this antibody to see if it specific for p190 RhoGAP A isoform. However, I checked the immunogen sequence an...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"