PathHunter® U2OS INSRa Functional Bioassay Kit

Background Information:

The PathHunter® U2OS INSRa Functional Bioassay Kit utilizes Enzyme Fragment Complementation (EFC) technology, where the β-galactosidase (β-gal) enzyme is split into two fragments, the ProLink™ (PK), and the Enzyme Acceptor (EA). Independently, these fragments have no β-gal enzymatic activity; however, when forced to complement through protein-protein interactions, they form an active β-gal enzyme.
The PathHunter INSRa Bioassay monitors activation of the INSRa receptor tyrosine kinase (RTK) in response to Insulin binding. In this system, INSRa is fused to the small β-gal fragment, PK, at the C-terminus of the receptor. The EA fragment of β-gal is fused to an SH2 domain-containing protein that recognizes a phosphotyrosine domain in the activated RTK. Ligand-induced activation by Insulin results in receptor phosphorylation. The SH2-EA fusion protein binds the phosphorylated receptor, forcing complementation of PK and EA to form an active β-gal enzyme. β-gal enzymatic activity is quantitatively measured using a chemiluminescent substrate provided in the PathHunter Detection Kit.
Bioassay kits are a convenient, ready-to-use format which contains all materials needed to run the assay, including single-use vials of cryopreserved cells, cell plating media, control agonist (in cluded in the 2-plate format), detection reagent, and assay plates.
Please reach out to Tech Support at DiscoverXSupportUS@discovery.eurofinsus.com for lot-specific CoA.

Target Class:

Kinase

Family:

RTK

Sub Family:

Protein Tyrosine

Protein Name:

IR

Protein Aliases:

CD antigen CD220 [Cleaved into: Insulin receptor subunit alpha; Insulin receptor subunit beta]

Accession Number:

NM_001079817

UniProt Number:

P06213

Gene Name:

INSR

Gene ID:

3643

Gene Aliases:

CD220

Target Species:

Human

Cell Background:

U2OS