Gα₁₃ Pull-Down Activation Assay Kit

Cat. # TD-80401

Background

A structurally diverse repertoire of ligands, from photons to large peptides, activates GPCRs to elicit their physiological functions. Ligand-bound GPCRs, in turn, function as guanine nucleotide exchange factors catalyzing the exchange of GDP bound on the Gα subunit with GTP in the presence of Gβγ, causing the dissociation of the Gα subunit from the Gβγ dime to form two functional units (Gα and Gβγ). Both Gα and Gβγ subunits signal to various cellular signaling pathways. Based on the sequence and functional homologies, G proteins are grouped into four families: G_s, G_i, GG_q, and G₁₂. As increasing numbers of effectors and interacting proteins for these G proteins have been identified, the physiological processes in which G proteins participate are multiplying.

Among the four subfamilies of G proteins, the function of G_12/13 subfamily is less well understood. In this family, there are two members, G₁₂ and G₁₃, that are expressed ubiquitously. Gα12 knockout mice appeared normal. Gα13 knockout mice displayed embryonic lethality (~E9.5). The Gα13-/- mouse embryos had defective vascular systems. G₁₃ is also essential for receptor tyrosine kinase-induced migration of fibroblast and endothelial cells.

Assay Principle

The Gα₁₃ Activation Assay Kit uses configuration-specific anti-Gα₁₃-GTP Mouse monoclonal antibody to measure Gα₁₃-GTP levels in cell extracts or in vitro GTPγS loading Gα₁₃ activation assays. Anti-Gα₁₃-GTP mouse monoclonal antibody is first incubated with cell lysates containing Gα₁₃-GTP. Next, the GTP-bound Gα₁₃ is pulled down by protein A/G agarose. Finally, the precipitated Gα₁₃-GTP is detected through immunoblot analysis using anti-Gα₁₃ mouse monoclonal antibody.