Anti-CDC42 antibody (ab64533)
Key features and details
- Rabbit polyclonal to CDC42
- Suitable for: ICC/IF, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-CDC42 antibody
See all CDC42 primary antibodies -
Description
Rabbit polyclonal to CDC42 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Pig, Saccharomyces cerevisiae, Chimpanzee, Zebrafish
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human CDC42.
Read Abcam's proprietary immunogen policy (Peptide available as ab66571.) -
General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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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 purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Positive Controls
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Recombinant Protein
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). Target
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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
modificationsAMPylation 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
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Database links
- Entrez Gene: 998 Human
- Entrez Gene: 12540 Mouse
- Entrez Gene: 780428 Pig
- Entrez Gene: 64465 Rat
- Omim: 116952 Human
- SwissProt: P60953 Human
- SwissProt: P60766 Mouse
- SwissProt: Q007T2 Pig
see all -
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
see all
Images
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Western blot - Anti-CDC42 antibody (ab64533)All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CDC42 knockout HEK-293T cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 21 kDaLanes 1-4: Merged signal (red and green). Green - ab64533 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab64533 Anti-CDC42 antibody was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab64533 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 µg/ml and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody (ab64533)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. -
Western blot - Anti-CDC42 antibody (ab64533)All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : CDC42 knockout HAP1 cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 21 kDaLanes 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 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CDC42 knockout samples were eexamined. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. Ab64533 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.
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Western blot - Anti-CDC42 antibody (ab64533)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
All lanes : 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 -
Western blot - Anti-CDC42 antibody (ab64533)All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml
Lane 1 :Recombinant Human CDC42 protein (ab87713) at 0.1 µg
Lane 2 :Recombinant Human CDC42 protein (ab87713) at 0.01 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 21 kDa
Exposure time: 1 minute
Protocols
Datasheets and documents
References (30)
ab64533 has been referenced in 30 publications.
- Sun P et al. A mosquito salivary protein promotes flavivirus transmission by activation of autophagy. Nat Commun 11:260 (2020). PubMed: 31937766
- Ma LL et al. Cdc42 subcellular relocation in response to VEGF/NRP1 engagement is associated with the poor prognosis of colorectal cancer. Cell Death Dis 11:171 (2020). PubMed: 32139668
- Meng J et al. Hydrogen gas represses the progression of lung cancer via down-regulating CD47. Biosci Rep 40:N/A (2020). PubMed: 32314789
- Woeste MA et al. Species-specific differences in nonlysosomal glucosylceramidase GBA2 function underlie locomotor dysfunction arising from loss-of-function mutations. J Biol Chem 294:3853-3871 (2019). PubMed: 30662006
- Mao F et al. miR-149 inhibits cell proliferation and enhances chemosensitivity by targeting CDC42 and BCL2 in neuroblastoma. Cancer Cell Int 19:357 (2019). PubMed: 31889909
Images
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Western blot - Anti-CDC42 antibody (ab64533)All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CDC42 knockout HEK-293T cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 21 kDaLanes 1-4: Merged signal (red and green). Green - ab64533 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab64533 Anti-CDC42 antibody was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab64533 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 µg/ml and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
-
Immunocytochemistry/ Immunofluorescence - Anti-CDC42 antibody (ab64533)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.
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Western blot - Anti-CDC42 antibody (ab64533)All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : CDC42 knockout HAP1 cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 21 kDaLanes 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 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CDC42 knockout samples were eexamined. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. Ab64533 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 (ab64533)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
All lanes : 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
-
Western blot - Anti-CDC42 antibody (ab64533)All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml
Lane 1 :Recombinant Human CDC42 protein (ab87713) at 0.1 µg
Lane 2 :Recombinant Human CDC42 protein (ab87713) at 0.01 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 21 kDa
Exposure time: 1 minute
