Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-CDC42 antibody [EPR15620] - Low endotoxin, Azide free (ab221781)

Overview

  • Product name

    Anti-CDC42 antibody [EPR15620] - Low endotoxin, Azide free
    See all CDC42 primary antibodies
  • Description

    Rabbit monoclonal [EPR15620] to CDC42 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, ICC/IF, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment 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

    ab221781 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab221781 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 at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).

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

  • 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 for detection at 1/1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187643).

  • 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). 

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187643).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187643).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187643).

  • 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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187643).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187643).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187643).

References

ab221781 has not yet been referenced specifically in any publications.

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