Overview

  • Product name
  • Description
    Rabbit polyclonal to CDC42
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Saccharomyces cerevisiae
    Predicted to work with: Pig, Chimpanzee, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human CDC42.

    (Peptide available as ab66571.)

  • Positive control
    • This antibody gave a positive signal in the following Human Whole Cell Lysates: HeLa, Jurkat, HepG2, MCF7, U2OS, A431 (data not shown), HEK 293 (data not shown) and SHSY-5Y (data not shown)

Properties

Applications

Our Abpromise guarantee covers the use of ab64533 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 a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
IHC-Fr 1/100.
IHC-P Use at an assay dependent concentration.

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

Anti-CDC42 antibody images



  • Predicted band size : 21 kDa

    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 – ab64533 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    Ab64533 was shown to recognize CDC42 when CDC42 knockout samples were used, along with additional cross-reactive bands. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. Ab64533 and ab8245 (loading control to GAPDH) were diluted at 1 µg/ml 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 h at room temperature before imaging.

  • All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 5 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Observed band size : 21 kDa
    Additional bands at : 52 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa


    Exposure time : 1 minute
  • ICC/IF image of ab64533 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64533, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive result in 4% pfa fixed (10 min) HeLa cells at 5µg/ml.

References for Anti-CDC42 antibody (ab64533)

This product has been referenced in:
  • Gerasimcik N  et al. The Rho GTPase Cdc42 Is Essential for the Activation and Function of Mature B Cells. J Immunol 194:4750-8 (2015). Read more (PubMed: 25870239) »
  • Zhao C  et al. microRNA-195 inhibits cell proliferation in bladder cancer via inhibition of cell division control protein 42 homolog/signal transducer and activator of transcription-3 signaling. Exp Ther Med 10:1103-1108 (2015). WB ; Human . Read more (PubMed: 26622447) »

See all 6 Publications for this product

Product Wall

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Zebrafish Tissue sections (retina)
Specification
retina
Permeabilization
Yes - triton X
Fixative
Paraformaldehyde
Username

Dr. Ryan Macdonald

Verified customer

Submitted May 22 2014

The nucleotide binding sites of this protein correspond to amino acids 10-17, 57-61 and 115-118.
Since the immunogen of this antibody corresponds to amino acids xxxxxxx, I can confirm that this antibody will not be able to distinguish between GTP ...

Read More

ab41429 does not detect RhoA, we have tested this directly using recombinant RhoA protein versus recombinant CDC42 protein. However, we have not tested reactivity to Rac1 and therefore, we cannot comment on the cross-reactivity with Rac1 since the epit...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Permeabilization
Yes - saponin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 17 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (SPhi992 strain)
Loading amount
50 µg
Specification
SPhi992 strain
Gel Running Conditions
Reduced Denaturing (12% Bis-TRIS in MOPS buffer)
Blocking step
Odyssey Blocking Agent (Lycor system) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MPRO (mouse promyelocytic) cells)
Loading amount
50 µg
Specification
MPRO (mouse promyelocytic) cells
Gel Running Conditions
Reduced Denaturing (12% Bis-TRIS in MOPS buffer)
Blocking step
Odyssey Blocking Agent (Lycor system) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Loading amount
50 µg
Specification
HEK293
Gel Running Conditions
Reduced Denaturing (12% Bis-TRIS in MOPS buffer)
Blocking step
Odyssey Blocking Agent (Lycor system) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 10 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (colon)
Specification
colon
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%
Username

Abcam user community

Verified customer

Submitted Feb 05 2010

Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa, Jurkat)
Loading amount
500 µg
Specification
HeLa, Jurkat
Gel Running Conditions
Reduced Denaturing (12,5% acrylamide gel)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
Username

Mrs. Merly Saare

Verified customer

Submitted Dec 07 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton x-100 in PBS
Blocking step
Milk as blocking agent for 40 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Mrs. Merly Saare

Verified customer

Submitted Sep 14 2009

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

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