Ran Pull-Down Activation Assay Kit

Cat. # TD-81701

Background

Small GTPases are a super-family of cellular signaling regulators. Ran (Ras-related nuclear protein) is a member of the Ras-superfamily GTPases. Ran is involved in the transport of proteins across the nuclear envelope, as well as in microtubule organization during mitosis. Currently there is no direct assay to measure the activation of Ran GTPases.

The Ran Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Ran-GTP, but not Ran-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.

Assay Principle

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

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