Antibodies, Proteins, Kits and Reagents for Life Science

Key features and details

Rabbit polyclonal to CDC42
Suitable for: ICC/IF, WB
Knockout validated
Reacts with: Human
Isotype: IgG

Product name

Anti-CDC42 antibody
See all CDC42 primary antibodies
Description

Rabbit polyclonal to CDC42
Host species

Rabbit
Tested Applications & Species

Application Species
ICC/IF
Human

WB
Human

See all applications and species data
Immunogen

Synthetic peptide corresponding to Human CDC42 aa 100 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available as ab66572)
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)

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS

Batches of this product that have a concentration
Concentration information loading...
Purity

Immunogen affinity purified
Clonality

Polyclonal
Isotype

IgG
Research areas

Compatible Secondaries
Isotype control
- Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
Positive Controls
- Recombinant Human CDC42 protein (ab87713)
Recombinant Protein
- Recombinant Human CDC42 protein (ab87713)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab66573 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
ICC/IF	Human
WB	Human
All applications	Mouse Chimpanzee Zebrafish

Application	Abreviews	Notes
ICC/IF		Use a concentration of 10 µg/ml.
WB		Use a concentration of 1 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).

Notes
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).

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.
Target information above from: UniProt accession P60953 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 998 Human
- Entrez Gene: 12540 Mouse
- Omim: 116952 Human
- SwissProt: P60953 Human
- SwissProt: P60766 Mouse
- Unigene: 467637 Human
- Unigene: 1022 Mouse
- Unigene: 447553 Mouse
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

Western blot - Anti-CDC42 antibody (ab66573)

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 - ab66573 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab66573 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. ab66573 and ab8245 (loading control to GAPDH) were diluted at 1µg/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 hour at room temperature before imaging.
Western blot - Anti-CDC42 antibody (ab66573)

All lanes : Anti-CDC42 antibody (ab66573) 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
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size: 21 kDa
Observed band size: 21 kDa
Additional bands at: 42 kDa, 48 kDa, 52 kDa. We are unsure as to the identity of these extra bands.

Exposure time: 30 seconds
Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody (ab66573)

ICC/IF image of ab66573 stained MCF7 cells. The cells were 4% PFA fixed (10 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 (ab66573, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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) at a concentration of 1.43µM.

Western blot protocols
Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheet

Publishing research using ab66573? Please let us know so that we can cite the reference in this datasheet.

ab66573 has been referenced in 1 publication.

Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371

Western blot - Anti-CDC42 antibody (ab66573)

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 - ab66573 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab66573 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. ab66573 and ab8245 (loading control to GAPDH) were diluted at 1µg/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 hour at room temperature before imaging.
Western blot - Anti-CDC42 antibody (ab66573)

All lanes : Anti-CDC42 antibody (ab66573) 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
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size: 21 kDa
Observed band size: 21 kDa
Additional bands at: 42 kDa, 48 kDa, 52 kDa. We are unsure as to the identity of these extra bands.

Exposure time: 30 seconds
Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody (ab66573)

ICC/IF image of ab66573 stained MCF7 cells. The cells were 4% PFA fixed (10 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 (ab66573, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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) at a concentration of 1.43µM.