RheB Pull-Down Activation Assay Kit

Cat. # TD-81201

Background

Small GTPases are a super-family of cellular signaling regulators. RheB is a member of the Ras-superfamily GTPases. RheB has been shown to interact with C-Raf, Mammalian target of rapamycin (mTOR), TSC2, Ataxia telangiectasia mutated (ATM), KIAA1303 and Ataxia telangiectasia and Rad3 related. Currently there is no direct assay to measure the activation of RheB GTPases.

The RheB Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes RheB-GTP, but not RheB-GDP. Given the high affinity of monoclonal antibodies to their antigens, the activation assay could be performed in a short time. This assay provides the reliable results with consistent reproducibility. These anti-RheB-GTP monoclonal antibodies can also be used to monitor the activation of RheB in cells and in tissues by immunohistochemistry.

Assay Principle

The RheB Activation Assay Kit uses configuration-specific anti-RheB-GTP Mouse monoclonal antibody to measure RheB-GTP levels in cell extracts or in vitro GTPγS loading RheB activation assays. Anti-RheB-GTP mouse monoclonal antibody is first incubated with cell lysates containing RheB-GTP. Next, the GTP-bound RheB is pulled down by protein A/G agarose. Finally, the precipitated RheB-GTP is detected through immunoblot analysis using Anti-RheB Rabbit Polyclonal Antibody.

The anti-RheB-GTP monoclonal antibody can also be used to monitor the activation of RheB in cells and in tissues by immunohistochemistry.