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Anti-Active Rac-GTP Mouse Monoclonal Antibody, 30 µl (26903)
Background Small GTPases are a superfamily of cellular signaling regulators. Rac belongs to the Rho sub-family of GTPases that regulate cell motility, cell division, and gene transcription. GTP binding increases the activity of Rac, and the hydrolysis of GTP to GDP renders it inactive.Currently the activation of Rac proteins is assayed with the binding of GTP-bound Rac to the p21-binding domain (PBD) of p21-activated protein kinase (PAK). This method is based on the observation that the active, GTP-bound Rac could bind to the PBD of PAK. However, the reproducibility of this method is poor. This is partially due to the relatively quick hydrolysis of GTP to GDP during the assay procedure, and the low binding affinity of PBD to Rac-GTP. Tested Applications IP, IHC Source Species Mouse Species Reactivity Anti-active Rac antibody recognizes active Rac1(maybe Rac2 and Rac3) from vertebrates. Datasheet Immunoprecipitation/Western Blot MEF cells were treated with (lane 2) or without (lane 1) PDGF. Cell lysates were incubated with an anti-active Rac monoclonal antibody (Cat. #NEBA10210) (top panel). The precipitated active Rac was immunoblotted with an anti-Rac rabbit polyclonal antibody (Cat. #NEBA10003). The bottom panel shows the Western blot with anti-Rac of the cell lysates used (5% of that used in the top panel). Immunohistochemistry Immunohistochemistry for the active Rac using Anti-Active Rac1-GTP Mouse Monoclonal Antibody (Cat. #NEBA10210) shows Rac-GTP immunolabeling (green) in combination with cofilin (red) on brain tissue sections. The tissue sections were fixed with -20 C methanol or 4% paraformaldehyde (fixation time 1hr) and stained with antibody at 1:1000 in 0.1M Phosphate buffer with 0.3% Triton X, and 4% BSA for 24h at room temperature. Secondary antibodies were anti-mouse AlexaFluor488 and anti-rabbit AlexaFluor594 at 1:1000.
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