Description
Description: This assay employs a two-site sandwich ELISA to quantitate DCUN1D5 in samples. An antibody specific for DCUN1D5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any DCUN1D5 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for DCUN1D5 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 DCUN1D5 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: DCUN1D5 Contains 1 DCUN1 domain.SCF-type E3 ubiquitin ligases are multi-protein complexes required for polyubiquitination and subsequent degradation of target proteins by the 26S proteasome.C. elegans DCN-1 and S. cerevisiae Dcn1p (defective in cullin neddylation) are characterized by a novel UBA-like ubiquitin-binding domain and a DUF298 domain of unknown function. Consistent with their requirements for neddylation, DCN-1 and Dcn1p directly bind Nedd8 and physically associate with cullins in both species. Moreover, overexpression of Dcn1p in yeast results in the accumulation of Nedd8-modified cullin Cdc53p. Both in vivo and in vitro experiments indicate that Dcn1p does not inhibit deneddylation of Cdc53p by the COP9 signalosome, but greatly increases the kinetics of the neddylation reaction