Description
This kit is designed for the detection of binding activity between EGF and EGFR using a homogeneous AlphaLISA™ assay (no wash steps). This assay can facilitate the design and development of therapetics by using competitive binding.
Features:
No-wash steps, no separation steps
Ease-of-use: few addition steps, fast assay development
Broad range of affinities: detect strong or weak interactions, from pM to mM affinity
Distance: measure very large protein or antibody complexes – spanning up to 200 nm or more
High avidity: multiple binding sites on each bead enables use of nanomolar concentrations of antibodies or proteins, as well as use of low affinity binders
AlphaLISA technology allows the detection of molecules of interest in buffer, cell culture media, serum and plasma in a highly sensitive, quantitative, reproducible and user-friendly mode. In this AlphaLISA assay, a biotinylated EGF binds to the Streptavidin-coated Alpha Donor beads, while EGFR-Fc is captured by Anti-Human IgG Fc-specific AlphaLISA Acceptor beads. When EGF is bound to EGFR, Donor beads and Acceptor beads come into close proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfers in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
Epidermal Growth Factor Receptor (EGFR, Her1, ErbB1) is a 134 kDa cell surface receptor, part of a four member subfamily of receptor tyrosine kinases (erbB1, erbB2, erbB3, and erbB4). EGFR binds to a family of proteins called the epidermal growth factors (primarily EGF), which, upon association induces dimerization of the receptor and initiates signal transduction for the promotion of cell survival and growth. Mutations that cause aberrant overexpression of EGF and/or EGFR have been linked with many different cancers such as non-small-cell lung and colon cancers. Drugs are being developed as a means to better treat these types cancer.
