Gαs,Gαs-GTP

Activation Assay Kit

Configuration-specific monoclonal antibody that recognizes Gαs-GTP, but not Gαs-GDP The only available Gαs activation assay kit directly measures the level of Gαs-GTP Providing reliable results with consistent reproducibility than other Gαs activation assay kits based on certain downstream signaling pathways Providing shorter experimental time Could be used to monitor the activation of Gαs in cells and in tissues by immunohistochemistry

Tanda Biosciences Gαs Activation Assay Kit bases on the configuration-specific anti-Gαs-GTP monoclonal antibody to measure the active Gαs-GTP levels, either from cell extracts or from in vitro GTPγS loading Gαs activation assays. Briefly, anti-active Gαs mouse monoclonal antibody will be incubated with cell lysates containing Gαs-GTP. The bound active Gαs will then be pulled down by protein A/G agarose. The precipitated active Gαs will be detected by immunoblot analysis using anti- Gαs rabbit polyclonal antibody.

A structurally diverse repertoire of ligands, from photons to large peptides, activates G protein-coupled receptors (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 dimer 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: Gs, Gi, Gq, and G12. Gαs family relays signals from many GPCRs to regualte various biological functions such as the stimulation of adenylyl cyclases. There were no direct methods to measure the activation of Gαs proteins by receptors (until this assay kit). Most reports used one downstream pathway, the increase of cAMP, as a readout. Tanda Biosciences Gαs Activation Assay Kit provides a direct measurement of the activation of Gαs proteins. This is a simple and fast tool to monitor the activation of Gαs. Each kit provides sufficient quantities to perform 30 assays. Tanda Biosciences Gαs Activation Assay Kit is based on the monoclonal antibody specifically recognizing the active GTP-bound Gαs proteins. This monoclonal antibody has much lower affinity towards the inactive Gαs proteins. Therefore, after activation by receptor signals, active GTP-bound Gαs proteins could be immunoprecipitated by this monoclonal antibody and further quantified by western blot with another anti- Gαs antibody.

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