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S08-
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Detailed Description
Small GTPases are a super-family of cellular signaling regulators. Ras belongs to the Ras sub-family of GTPases that regulate cell growth, cell motility, and gene transcription. GTP binding increases the activity of Ras, and the hydrolysis of GTP to GDP renders it inactive.
Currently the activation of Ras proteins is assayed with the binding of GTP-bound Ras to the Ras-binding domain (RBD) of Raf protein kinase. This method is based on the observation that the active, GTP-bound Ras could bind to the RBD of Raf. 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 RBD to Ras-GTP.
NewEast Biosciences Ras Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Ras-GTP, but not Ras-GDP. Given the high affinity of monoclonal antibodies to their antigens, the activation assay could be performed in a much shorter time. This assay provides the reliable results with consistent reproducibility.
These anti-Ras-GTP monoclonal antibody can also be used to monitor the activation of Ras in cells and in tissues by immunohistochemistry.
NewEast Biosciences Ras Activation Assay Kit provides a simple and fast method to monitor the activation of Ras. Each kit provides sufficient quantities to perform 20 assays.
Assay Principle
NewEast Biosciences Ras Activation Assay Kit bases on the configuration-specific anti-Ras-GTP monoclonal antibody to measure the active Ras-GTP levels, either from cell extracts or from in vitro GTP?S loading Ras activation assays. Briefly, anti-active Ras mouse monoclonal antibody will be incubated with cell lysates containing Ras-GTP. The bound active Ras will then be pulled down by protein A/G agarose. The precipitated active Ras will be detected by immunoblot analysis using anti-Ras mouse monoclonal antibody.
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