Description
This assay employs a two-site sandwich ELISA to quantitate EPS15 in samples. An antibody specific for EPS15 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyEPS15 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for EPS15 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of EPS15 bound in the initial step. The color development is stopped and the intensity of the color is measured.
EPS15 encodes a protein that is part of the EGFR pathway. The protein is present at clathrin-coated pits and is involved in receptor-mediated endocytosis of EGF. Notably, this gene is rearranged with the HRX/ALL/MLL gene in acute myelogeneous leukemias. Alternate transcriptional splice variants of this gene have been observed but have not been thoroughly characterized.Binding of the parkin ubiquitin-like domain to the Eps15 ubiquitin-interacting motifs was required for parkin-mediated Eps15 ubiquitination. Egfr endocytosis and degradation were accelerated in parkin-deficient cells, and Egfr signaling via the PI3K-Akt pathway was reduced in parkin-knockout mouse brain