Introduction
Rac1 is one of most extensively studied member of Rho GTPase family. This family is involved in a wide range of cellular responses, including cytoskeletal reorganization, regulation of transcription, cell migration, cellular transformation and metastasis. Rac1 is expressed in a large number of different cell types.
Product Uses Include
This antibody is recommended for detection of Rac1 in human, mouse, rat or other extracts. The following protocols have been tested with this antibody:
- Western blot: recommended
- Immunocytochemistry: recommended
Material
The anti-Rac1 antibody (Cat. # ARC03) is a mouse monoclonal antibody that only recognizes Rac1, not Rac2, Rac3, Cdc42 or other small GTPases. Other commercially available Rac1 antibodies recognize Rac2 and 3, some even recognize Cdc42 (Table 1 and Figure 1). Platelet extract (Cat. # EXT01) is included as a positive control. ARC03 is supplied as a lyophilized white powder.
Figure 1 legend - Western blot of purified Rac1, 2, 3, Cdc42 and platelet extracts probed with anti-Rac1 monoclonal antibody (Cat. # ARC03) and other commercially availabe Rac1 antibodies. The detection of 25 ng His-Rac1 ( lane 1), His-Rac2 (lane 2), GST-Rac3 (lane 3), GST-Cdc42 (lane 4) and 50 µg of platelet extract (lane 5). ARC03 does not cross-react with Rac2, 3 or Cdc42, while all other commercially available Rac1 antibody crossreact with GTPases other than Rac1. The blot was probed with a 1 µg/ml (1:500) dilution of ARC03. Other blots were probed according to manufacturers’ protocols.
Table 1 - Reactivity of other commercially available Rac antibodies
Rac1 | Rac2 | Rac3 | Cdc42 | |
Cytoskeleton, Inc. (Cat. # ARC03) | Yes | No | No | No |
BD Biosciences (Cat. #610650) | Yes | Yes | Yes | Yes |
Santa Cruz (Cat. #sc-95) | Yes | No | Yes | Yes |
Upstate (Cat. # 05-389) | Yes | No | Yes | No |
Immunocytochemistry
Figure 2 - Immunofluorescence image of mouse Swiss 3T3 cells stained with Anti-Rac1 monoclonal antibody (Cat. # ARC03). Swiss 3T3 cells were grown to semi-confluency and fixed with methanol. Immunofluorescence staining using 1 µg/ml (1:20 dilution) of ARC03 antibody is shown (red). The primary antibody was detected with a 1:500 dilution of goat anti-mouse rhodamine conjugated antibody. Photograph was taken with a 100X objective lens.
Other associated products:
Anti-RhoA monoclonal antibody (Cat.# ARH03)
Anti-Cdc42 monoclonal antibody (Cat.# ACD03)
Cdc42 G-LISA™ Activation Assay (Cat.# BK127)
RhoA G-LISA™ Activation Assay (Cat.# BK124)
Rac1 G-LISA™ Activation Assay (Cat.# BK126)
Rac1,2,3 G-LISA™ Activation Assay (Cat.# BK125)
For product Datasheets and MSDSs please click on the PDF links below. For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com
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Guo, Y. et al. Cytotoxic necrotizing factor 1 promotes bladder cancer angiogenesis through activating RhoC. FASEB J. 34, 7927–7940 (2020).
Am. J. Physiol. Gastrointest. Liver Physiol. 2015; 308, G92-G99."Cellular inhibitor of apoptosis protein 2 controls human colonic epithelial restitution, migration, and Rac1 activation"Author(s): Seidelin J.B. et al.(See PubMed article)
Integr Biol (Camb). 2014; 6, 885-94."Simultaneous and independent tuning of RhoA and Rac1 activity with orthogonally inducible promoters"Author(s): MacKay J.L. and Kumar S.(See PubMed article)
Immunol Cell Biol. 2014; 92, 275-86."The Rho GTPase Rac1 is required for recycling endosome-mediated secretion of TNF in macrophages"Author(s): Stanley A.C. et al.(See PubMed article)
Mol. Cell Biol. 2013; doi:10.1128/MCB.00822-13."Phosphorylation of Rac1 T108 by ERK in response to EGF: 1 A novel mechanism to regulate Rac1 function. "Author(s): Tong J., et al.(See PubMed article)
Cereb. Cortex. 2012; 22, 680-692."Rac1-dependent cell cycle exit of MGE precursors and GABAergic interneuron migration to the cortex. "Author(s): Vidaki et al.(See PubMed article)
Am. J. Pathol. 2012; 180, 526–540."Rac1b increases with progressive tau pathology within cholinergic nucleus basalis neurons in Alzheimer"s disease. "Author(s): Perez S.E., et al.(See PubMed article)
Neoplasia. 2012; 14, 84–94."Merlin/NF2 Regulates Angiogenesis in Schwannomas through a Rac1/Semaphorin 3F-Dependent Mechanism. "Author(s): Wong H.-K., et al.(See PubMed article)
J. Pathol. 2012; 226, 674-686."15-Hydroxyprostaglandin dehydrogenase associates with poor prognosis in breast cancer, induces epithelial–mesenchymal transition, and promotes cell migration in cultured breast cancer cells."Author(s): Lehtinen L., et al.(See PubMed article)
Question 1: What is the antigen that this antibody was raised against?
Answer 1: The Rac1 specific antibody (Cat. # ARC03) is a mouse monoclonal antibody and it was raised against the C-terminal peptide of human Rac1.
Question 2: What species has this antibody been tested against?
Answer 2: The Rac1 specific antibody has been used to label Rac1 proteins in human, bovine, rat and mouse extracts.
If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com.
