Validated using a knockout cell line
Recombinant
RabMAb

Anti-CDC42 antibody [EPR15620] (ab187643)

Overview

  • Product name
    Anti-CDC42 antibody [EPR15620]
    See all CDC42 primary antibodies
  • Description
    Rabbit monoclonal [EPR15620] to CDC42
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, IP, Flow Cyt, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human CDC42 aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: P60953

  • Positive control
    • HT-29, Jurkat and HeLa cell lysates; Mouse brain, Mouse kidney, Mouse spleen and Rat spleen lysates; Human breast carcinoma tissue; HeLa cells.
  • General notes

     

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab187643 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 1/250.

Perform heat mediated antigen retrieval using EDTA Buffer, pH9 before commencing with IHC staining protocol.

IP 1/20 - 1/40.
Flow Cyt 1/170.

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

WB 1/10000 - 1/50000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
ICC/IF 1/500.

Target

  • Function
    Plasma membrane-associated small GTPase which cycles between an active GTP-bound and an inactive GDP-bound state. In active state binds to a variety of effector proteins to regulate cellular responses. Involved in epithelial cell polarization processes. Causes the formation of thin, actin-rich surface projections called filopodia.
  • Sequence similarities
    Belongs to the small GTPase superfamily. Rho family. CDC42 subfamily.
  • Post-translational
    modifications
    AMPylation at Tyr-32 and Thr-35 are mediated by bacterial enzymes in case of infection by H.somnus and V.parahaemolyticus, respectively. AMPylation occurs in the effector region and leads to inactivation of the GTPase activity by preventing the interaction with downstream effectors, thereby inhibiting actin assembly in infected cells. It is unclear whether some human enzyme mediates AMPylation; FICD has such ability in vitro but additional experiments remain to be done to confirm results in vivo.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Form
    There are 2 isoforms produced by alternative splicing. Isoform 1 also known as: Brain; Isoform 2 also known as: Placental.
  • Alternative names
    • CDC42 antibody
    • CDC42_HUMAN antibody
    • CDC42Hs antibody
    • Cell division control protein 42 homolog antibody
    • Cell division cycle 42 (GTP binding protein 25kDa) antibody
    • Cell division cycle 42 antibody
    • dJ224A6.1.1 (cell division cycle 42 (GTP-binding protein, 25kD)) antibody
    • dJ224A6.1.2 (cell division cycle 42 (GTP-binding protein, 25kD)) antibody
    • G25K antibody
    • G25K GTP-binding protein antibody
    • Growth regulating protein antibody
    • GTP binding protein 25kDa antibody
    • Small GTP binding protein CDC42 antibody
    • TKS antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of U937 (Human histiocytic lymphoma cell line) labelling CDC42 with purified ab187643 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: CDC42 knockout HAP1 cell lysate (20 µg)
    Lane 3: MCF7 cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab187643 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab187643 was shown to specifically react with CDC42 in wild-type HAP1 cells. No band was observed when CDC42 knockout samples were examined. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab187643 and ab8245 (loading control to GAPDH) were diluted at 1/5000 and 1/10,000 respectively 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 hour at room temperature before imaging.

  • Lane 1 (input): HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate,10μg
    Lane 2(+): HT-29 whole cell lysate
    Lane 3(-): Rabbit monoclonal IgG (ab172730) instead of  ab187643 in HT-29 whole cell lysate

    Ab187643 immunoprecipitating CDC42 in HT-29 whole cell lysate. Capture antibody was used at a 1:60 dilution. For western blotting, ab187643 was used as the primary antibody at a 1:1000 dilution. Ab131366 Veriblot for IP (HRP) was used as the secondary antibody at 1:1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST

  • Flow cytometry analysis of HeLa cells (human cervix adenocarcinoma epithelial).  Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Primary antibody was used at a 1:120 dilution (red).  A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (black). Cell without incubation with primary antibody and secondary antibody (Blue). 

     

  • ab187643 staining CDC42 in Human lung carcinoma tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin-embedded sections). Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0).  Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on human lung carcinoma.

     

  • ab187643 staining CDC42 in Mouse colon tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin-embedded sections).  Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0).  Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on mouse colon.

  • ab187643 staining CDC42 in Rat colon tissue sections by Immunohistochemistry (IHC-P- paraformaldehyde-fixed, paraffin-embedded sections).  Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0).  Samples were incubated with primary antibody at a 1/500 dilution. A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Cytoplasmic staining on rat colon.

  • All lanes : Anti-CDC42 antibody [EPR15620] (ab187643) at 1/20000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HT-29 cell lysate
    Lane 3 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 21 kDa
    Observed band size: 21 kDa

  • All lanes : Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 21 kDa
    Observed band size: 21 kDa

  • Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling CDC42 with ab187643 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

References

This product has been referenced in:
  • Li M  et al. Total alkaloids ofCorydalis saxicolabunting inhibits migration of A549 cells by suppressing Cdc42 or Vav1. Oncol Lett 15:475-482 (2018). Read more (PubMed: 29285198) »
  • Peng H  et al. Qihuang decoction promotes the recovery of intestinal immune barrier dysfunction after gastrectomy in rats. Am J Transl Res 10:827-836 (2018). WB ; Rat . Read more (PubMed: 29636872) »

See all 10 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (3T3)
Permeabilization
Yes - triton X100
Specification
3T3
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Peter Gould

Verified customer

Submitted Nov 08 2016

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

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