Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate DLGAP3 in samples. An antibody specific for DLGAP3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyDLGAP3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for DLGAP3 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 DLGAP3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Using a segment of rat Fak (PTK2) to screen an adult human brain cDNA library, Bongiorno-Borbone et al. (2005) cloned DLGAP3, which they called SAPAP3. The deduced protein contains 977 amino acids. By Northern blot analysis, Takeuchi et al. (1997) found brain-specific expression of rat Sapap3. The rat protein localized to dendrites and cell bodies and was enriched in synaptosomal membrane fractions.By mutation analysis, Bongiorno-Borbone et al. (2005) demonstrated that the N-terminal half of human SAPAP3 interacted with a C-terminal segment of rat Fak. They also found that SAPAP3 interacted with Pyk2 (PTK2B) in in vitro pull-down assays. In rat forebrain lysates, part of Fak and Pyk2 had a similar subcellular distribution as Sapap3 and Sap90/Psd95, which supported their functional association.
