Small GTPases are a super-family 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.
These anti-Rac-GTP monoclonal antibody can also be used to monitor the activation of Rac in cells and in tissues by immunohistochemistry.
NewEast Biosciences Rac Activation Assay Kit provides a simple and fast method to monitor the activation of Rac. Each kit provides sufficient quantities to perform 20 assays.
Assay Principle
NewEast Biosciences Rac Activation Assay Kit bases on the configuration-specific anti-Rac-GTP monoclonal antibody to measure the active Rac-GTP levels, either from cell extracts or from in vitro GTPγS loading Rac activation assays. Briefly, anti-active Rac mouse monoclonal antibody will be incubated with cell lysates containing Rac-GTP. The bound active Rac will then be pulled down by protein A/G agarose. The precipitated active Rac will be detected by immunoblot analysis using anti-Rac rabbit polyclonal antibody.
Cart
Product Manual