Description
Principle of the assay: The microtiter plate provided in this kit has been pre-coated with biotin-BSA and Avidin. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated GPR101 antibody and incubated. Then Horseradish Peroxidase (HRP)-conjugated antibody preparation specific for GPR101 are added and incubated. Substrate solutions are added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm±2 nm. The concentration of GPR101 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Background: G protein-coupled receptors (GPCRs, or GPRs) contain 7 transmembrane domains and transduce extracellular signals through heterotrimeric G proteins.Lee et al. (2001) identified GPR101 within a genomic database using sequences of the histamine receptor H1 as query. PCR primers were designed to amplify and clone GPR101 from a genomic library. Full-length GPR101 encodes a deduced 508-amino acid protein that shares approximately 30% sequence identity in the transmembrane regions with the G protein-coupled receptor RE2, the serotonin 5HT1A receptor (HTR1A), and the alpha-1A-adrenergic receptor. Northern blot analysis of human brain tissues revealed expression of 9.5- and 4.2-kb transcripts in the caudate putamen and hypothalamus. No expression was detected in cortex, thalamus, hippocampus and pons