Arl1 Pull-Down Activation Assay Kit

Cat. # TD-82901

Background

Arf-like protein 1 (Arl1) is a member of the Arf family of regulatory GTPases, within the Ras superfamily of GTPases, and with highly conserved orthologs throughout eukaryotes. Arl1 is essential for early embryonic development in Drosophila and in Caenorhabditis elegans. Arl1 is most similar in primary sequence, cellular location, and function (regulation of membrane traffic) to Arf1–6 and even shares several common binding partners. In addition to its function in membrane traffic at the Golgi/trans-Golgi network, there are reports indicating a possible role for Arl1 in ion homeostasis in yeast.

Currently there is no direct assay to measure the activation of Arl1 GTPases.

The Arl1 Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Arl1-GTP, but not Arl1-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 Arl1 Activation Assay Kit uses configuration-specific anti-Arl1-GTP Mouse monoclonal antibody to measure Arl1-GTP levels in cell extracts or in vitro GTPγS loading Arl1 activation assays. Anti-Arl1-GTP mouse monoclonal antibody is first incubated with cell lysates containing Arl1-GTP. Next, the GTP-bound Arl1 is pulled down by protein A/G agarose. Finally, the precipitated Arl1-GTP is detected through immunoblot analysis using anti-Arl1 mouse monoclonal antibody.