Description
Description: This assay employs a two-site sandwich ELISA to quantitate ERO1L in samples. An antibody specific for ERO1L has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ERO1L present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for ERO1L 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 ERO1L bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: ERO1-L shares extensive homology with the Saccharomyces cerevisiae ERO1 gene, required in yeast for oxidative protein folding. When expressed in mammalian cells, the product of the human ERO1-L gene co-localizes with ER markers and displays Endo-H-sensitive glycans. In isolated microsomes, ERO1-L behaves as a type II integral membrane protein. ERO1-L is able to complement several phenotypic traits of the yeast thermosensitive mutant ero1-1, including temperature and dithiothreitol sensitivity, and intrachain disulfide bond formation in carboxypeptidase Y. ERO1-L is no longer functional when either one of the highly conserved Cys-394 or Cys-397 is mutated. These results strongly suggest that ERO1-L is involved in oxidative ER protein folding in mammalian cells