Sar1 Pull-Down Activation Assay Kit

Cat. # TD-81801

Background

Small GTPases are a super-family of cellular signaling regulators. Sar1 is a member of these small GTPases. Sar1 functions as a molecular switch to control protein–protein and protein–lipid interactions that direct vesicle budding from the ER. Sar1 is involved in membrane trafficking, and regulates the assembly and disassembly of COPII coats. Currently there is no direct assay to measure the activation of Sar1 GTPases.

The Sar1 Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Sar1-GTP, but not Sar1-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 Sar1 Activation Assay Kit uses configuration-specific anti-Sar1-GTP Mouse monoclonal antibody to measure Sar1-GTP levels in cell extracts or in vitro GTPγS loading Sar1 activation assays. Anti-Sar1-GTP mouse monoclonal antibody is first incubated with cell lysates containing Sar1-GTP. Next, the GTP-bound Sar1 is pulled down by protein A/G agarose. Finally, the precipitated Sar1-GTP is detected through immunoblot analysis using Anti-Sar1 Rabbit Polyclonal Antibody.

The anti-Sar1-GTP monoclonal antibody can also be used to monitor the activation of Sar1 in cells and in tissues by immunohistochemistry.