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- 產(chǎn)品名稱： Gαo Activation Assay Kit
- 產(chǎn)品貨號： 80901
- 貨期： 現(xiàn)貨
- 價格與訂購： 6800
- 數(shù)量：
庫存： 5
- 規(guī)格： 96T
- 產(chǎn)品信息
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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.
Bioyears 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.
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Assay Principle
Bioyearst 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.
Kit Components
1. Anti active Gαo, Mouse Monoclonal Antibody (Catalog No. 26907): One vial – 22 μL (1mg/mL) in
PBS, pH 7.4, contained 50% glycerol. This antibody specifically recognizes GTP- Gαo from all ertebrates.
2. Protein A/G Agarose (Catalog No. 30301): One vial – 400 μL of 50% slurry.
3. 5X Assay/Lysis Buffer (Catalog No. 30303): One bottle – 30 mL of 250 mM Tris-HCl, pH 7.4,750 mM
NaCl, 5 mM EDTA, 5% Triton X-100.
4. Anti Gα0, Rabbit Polyclonal Antibody (Catalog No. 21015): One vial – 22 μL(1 mg/mL) in PBS, pH 7.4,
contained 50% glycerol.
5. 100 X GTPγS (Catalog No. 30302): One vial –100 μL at 10 mM, use 5 μL of GTPγS for GTP-labeling of
0.5 mL of cell lysate.
6. 100 X GDP (Catalog No. 30304): One vial –100 μL at 100 mM, use 5 μL of GDP for GDP-labeling of 0.5
mL of cell lysate.
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Storage
Store all kit components at 4oC until their expiration dates. For long-term usage (more than 6 months),
please keep the antibodies at -20oC.
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Note
For Research Use Only.
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