Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PHACTR1 in samples. An antibody specific for PHACTR1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPHACTR1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PHACTR1 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 PHACTR1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:The PHACTR proteins share highest similarity in sequences surrounding the N- and C-terminal RPEL repeats and in the C-terminal actin- and PP1-binding domains. In situ hybridization of rat brain showed enrichment of Phactr1 in cerebral cortex, hippocampus, olfactory tubercle, nucleus accumbens, caudate-putamen, and piriform cortex.
Endogenous PP1 coprecipitated with ectopically expressed rat Phactr1 in human embryonic kidney cells. Phactr1 also coprecipitated with actin in rat brain lysates. Yeast 2-hybrid analysis of a series of Phactr1 C-terminal truncation mutants indicated that PP1 bound to Phactr1 near the C terminus, while actin bound in a region containing RPEL repeats 2 and 3.
