Rap1 Pull-Down Activation Assay Kit

Cat. # TD-81401

Background

Small GTPases are a super-family of cellular signaling regulators. The small Ras-like GTPase Rap1 is an evolutionary conserved protein that originally gained interest because of its capacity to revert the morphological phenotype of Ras-transformed fibroblasts. Rap1 is regulated by a large number of stimuli that include growth factors and cytokines, but also physical force and osmotic stress. Rap1 was shown to regulate multiple basic cellular processes. The best studied aspect of Rap1 function in endothelial cells involved its role in regulation of cell-cell junction formation and remodeling.

Rap1 Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Rap1-GTP, but not Rap1-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.

The anti-Rap1-GTP monoclonal antibody can also be used to monitor the activation of Rap1 in cells and in tissues by immunohistochemistry.

Assay Principle

The Rap1 Activation Assay Kit uses configuration-specific anti-Rap1-GTP Mouse monoclonal antibody to measure Rap1-GTP levels in cell extracts or in vitro GTPγS loading Rap1 activation assays. Anti-Rap1-GTP mouse monoclonal antibody is first incubated with cell lysates containing Rap1-GTP. Next, the GTP-bound Rap1 is pulled down by protein A/G agarose. Finally, the precipitated Rap1-GTP is detected through immunoblot analysis using Anti-Rap1 Rabbit Polyclonal Antibody.