Rab5B Pull-Down Activation Assay Kit

Cat. # TD-83401

Background

Small GTPases are a super-family of cellular signaling regulators. Rab5 is a member of the Rab-family GTPases. Rab5 is a rate-limiting component of the machinery regulating the kinetics of membrane traffic in the early endocytic pathway.

Currently there is no direct assay to measure the activation of Rab5B GTPases.

Rab5B Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Rab5-GTP, but not Rab5-GDP, and a Rab5B specific rabbit polyclonal antibody. 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 Rab5B Activation Assay Kit uses configuration-specific anti-Rab5-GTP Mouse monoclonal antibody to measure Rab5-GTP levels in cell extracts or in vitro GTPγS loading Rab5 activation assays. Anti-Rab5-GTP mouse monoclonal antibody is first incubated with cell lysates containing Rab5-GTP. Next, the GTP-bound Rab5 is pulled down by protein A/G agarose. Finally, the precipitated Rab5-GTP is detected through immunoblot analysis using Anti-Rab5B Rabbit Polyclonal Antibody.