Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate RAPGEF4 in samples. An antibody specific for RAPGEF4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyRAPGEF4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RAPGEF4 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 RAPGEF4 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Sequence analysis predicted that the 1,011-amino acid cAMP-GEFII protein contains an N-terminal cAMP-binding domain and a C-terminal GEF domain. Expression of cAMP-GEFI and cAMP-GEFII activated RAP1A after forskolin and 3-isobutyl-1-methylxanthine stimulation, independent of the protein kinase A pathway.
cAMP-GEFI and cAMP-GEFII expression did not activate or only slightly activated other RAS superfamily members after stimulation. Northern blot analysis detected expression of a 4.4-kb cAMP-GEFII transcript that was most prominent in brain and adrenal gland. Within the brain, expression was strongest in cortex, occipital pole, frontal lobe, temporal lobe, amygdala, putamen, hippocampus, and cerebellum.
