RalA Pull-Down Activation Assay Kit

Cat. # TD-83601

Background

Small GTPases are a super-family of cellular signaling regulators. The Ras-like small G proteins, RalA/B, are important components of Ras signaling pathways, implicated in the initiation and maintenance of tumorigenic transformation, as well as vesicle transport, apoptosis, transcription, cell migration, and cell proliferation.

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

The RalA Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Ral-GTP, but not Ral-GDP, and a RalA 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.

These anti-Ral-GTP monoclonal antibodies can also be used to monitor the activation of Ral in cells and in tissues by immunohistochemistry.

Assay Principle

The RalA Activation Assay Kit uses configuration-specific anti-Ral-GTP Mouse monoclonal antibody to measure Ral-GTP levels either from cell extracts or from in vitro GTPγS loading Ral activation assays. Anti-Ral-GTP mouse monoclonal antibody is first incubated with cell lysates containing Ral-GTP. Next, the GTP-bound Ral is pulled down by protein A/G agarose. Finally, the precipitated active RalA or RalB will be detected by immunoblot analysis using anti- RalA or RalB rabbit polyclonal antibody, respectively.