Configuration-specific monoclonal antibody that recognizes Gαz-GTP, but not Gαz-GDP
The only available Gαz activation assay kit directly measures the level of Gαz-GTP
Providing reliable results with consistent reproducibility than other Gαz activation assay kits based on certain downstream signaling pathways
Providing shorter experimental time
Could be used to monitor the activation of Gαz in cells and in tissues by immunohistochemistry
Product Description
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 Ga subunit with GTP in the presence of Gβγ, causing the dissociation of the Ga subunit from the Gβγ dimer to form two functional units (Ga and Gβγ). Both Ga 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.
Gαi family (including Gαz) is the largest family of G proteins. They relay signals from many GPCRs to regualte various biological functions. There were no direct methods to measure the activation of Gαz proteins by receptors (until this assay kit). Most reports used one of the downstream pathway, i.e. the inhibition of adenylyl cyclases, as a readout.
NewEast Biosciences Gαz Activation Assay Kit provides a direct measurement of the activation of Gαz proteins. This is a simple and fast tool to monitor the activation of Gαz. Each kit provides sufficient quantities to perform 20 assays.
NewEast Biosciences Gαz Activation Assay Kit is based on the monoclonal antibody specifically recognizing the active GTP-bound Gαz proteins. This monoclonal antibody has much lower affinity towards the inactive Gαz proteins. Therefore, after activation by receptor signals, active GTP-bound Gαz proteins could be immunoprecipitated by this monoclonal antibody and further quantified by western blot with another anti- Gαz antibody.
Assay Principle
NewEast Biosciences Gαz Activation Assay Kit is an immunoprecipitation/western blot assay to measure the levels of active GTP-bound Gαz proteins, either from cell extracts or from in vitro GTPγS loaded Gαzproteins. Briefly, the anti-active Gαz monoclonal antibody will specifically bind to active Gαz protein. This antibody/Gαz complex will then be pulled down by protein A/G agarose. The precipitated active Gαz proteins will be detected by immunoblots with another anti-Gαz antibody.
