Technical Data
R0400
Rac1 (Ras-related C3 Botulinum Toxin Substrate 1 (Rho Family, Small GTP Binding Protein Rac1), Cell Migration-inducing Gene 5 Protein, MGC111543, MIG5, p21-Rac1, Ras-like Protein TC25, Ras-related C3 Botulinum Toxin Substrate 1, TC25, TC-25)
Description:
Ras related C3 botulinum toxin substrate 1 (RAC1) protein is a GTPase which belongs to the RAS superfamily of small GTP binding proteins. Members of this superfamily appear to regulate a diverse array of cellular events, including the control of cell growth, cytoskeletal reorganization, and the activation of protein kinases.

Applications:
Suitable for use in Western Blot, Immunohistochemistry, Affinity Precipitation and Immunoprecipitation. Other applications not tested.

Recommended Dilutions:
Western Blot: 0.5-2ug/ml. 20ug of rat brain microsomal prep was resolved by electrophoresis, transferred to PVDF or nitrocellulose and probed with R0400 (0.5ug/ml). Proteins were visualized using an IgG (HRP) goat anti-mouse and a chemiluminescence detection system.
Immunohistochemistry: 10ug/ml. Gave positive immunostaining of rat brain sections fixed first with PBS, 4% paraformaldehyde, 2% acetic acid for 30 minutes followed by ethanol:acetic acid [95:5] for 2 minutes.
Affinity Precipitation Assay: 5-10ug of R0400-55A PAK-1 PBD agarose (as part of R0400-55 Rac1 Activation Assay Kit, BioAssay™) precipitated Rac1-GTP in lysates of 3T3 fibroblasts that were loaded in vitro with GTP-g-S. Proteins were detected by Western Blot anaylsis using 1ug/ml.
Immunoprecipitation: 4ug immunoprecipitated Rac1 from a rat brain microsomal protein preparation.
Optimal dilutions to be determined by the researcher.

Recommended Secondary Antibody (WB):
I1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt x Mo
I1904-06H: IgG, H&L (HRP) (X-Adsorbed) Pab Gt x Mo
Recommended Secondary Antibodies (IHC):
I1904-06F: IgG, H&L (FITC) (X-Adsorbed) Pab Gt x Mo

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgG2b0.T.127Affinity Purified
SizeStorageShippingSourceHost
250ug-20°CBlue IceHumanMouse
Concentration:
~1mg/ml
Immunogen:
Recombinant protein containing the full length human Rac1 (Accession #: NP_008839).
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol.
Specificity:
Recognizes human Rac1 at ~21kD. Slight Rac2 crossreactivity is observed. An additional unknown protein of ~55kD may be detected in some preparations. Species Crossreactivity: mouse and rat
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
US Biological application reference: Gronborg, M. et al., (2006) Molecular and Cellular Proteomics, 5; 157-171. 1. Kjoller, L., and Hall, A. J. Cell Bio.. 152: 1145-1157 (2001). 2. Diakonova, M., et al., J. Biol. Chem. 277: 10,669-10,677 (2002). 3. Kovacic, H., et al., J. Biol. Chem. 276: 45,856-45,861 (2001). 4. Castoria, G., et al., J. Cell. Biol. 161: 547-556 (2003). 5. Gallager, B.C., et al., J. Cell. Sci. 116: 3239-3250 (2003). 6. Gismondi, M., et al., J. Immunol. 170: 3065-3073 (2003).