Rab35 Pull-Down Activation Assay Kit

Cat. # TD-82801

Background

The Rab family of GTPases regulates eukaryotic vesicular membrane traffic. Rab35 is with both plasma membrane and endosomal localization and has been implicated in diverse processes that include T-cell receptor recycling, oocyte yolk protein recycling and cytokinesis. Rab35 regulates neurite outgrowth in neuronal-like cells and can induce protrusions.

Currently there is no direct assay to measure the activation of Rab35 GTPases.

The Rab35 Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Rab35-GTP, but not Rab35-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 Rab35 Activation Assay Kit uses configuration-specific anti-Rab35-GTP Mouse monoclonal antibody to measure Rab35-GTP levels in cell extracts or in vitro GTPγS loading Rab35 activation assays. Anti-Rab35-GTP mouse monoclonal antibody is first incubated with cell lysates containing Rab35-GTP. Next, the GTP-bound Rab35 is pulled down by protein A/G agarose. Finally, the precipitated Rab35-GTP is detected through immunoblot analysis using Anti-Rab35 Rabbit Polyclonal Antibody.