Configuration-specific Monoclonal Antibody Based
Gαo Activation Assay Kit
Catalog Number： 80901
20 assays

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 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.
Gαi family （including Gαo） 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
Gαo 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 Gαo Activation Assay Kit provides a direct measurement of the activation of Gαo
proteins. This is a simple and fast tool to monitor the activation of Gαo. Each kit provides sufficient
quantities to perform 20 assays.
NewEast Biosciences Gαo Activation Assay Kit is based on the monoclonal antibody specifically
recognizing the active GTP-bound Gαo proteins. This monoclonal antibody has much lower affinity
towards the inactive Gαo proteins. Therefore, after activation by receptor signals, active GTP-bound
Gαo proteins could be immunoprecipitated by this monoclonal antibody and further quantified by
western blot with another anti- Gαo antibody.
Assay Principle
NewEast Biosciences Gαo Activation Assay Kit is an immunoprecipitation/western blot assay to
measure the levels of active GTP-bound Gαo proteins, either from cell extracts or from in vitro GTPγS
loaded Gαo proteins. Briefly, the anti-active Gαo monoclonal antibody will specifically bind to active
Gαo protein. This antibody/Gαo complex will then be pulled down by protein A/G agarose. The
precipitated active Gαo proteins will be detected by immunoblots with another anti-Gαo antibody.