Agouti-related protein, also called Agouti-related peptide (AgRP), is a neuropeptide produced in the brain by the AgRP/NPY neuron. It is only synthesised in NPY-containing cell bodies located in the ventromedial part of the arcuate nucleus in the hypothalamus. In humans, the agouti-related peptide is encoded by the AGRP gene. AgRP is co-expressed with Neuropeptide Y and works by increasing appetite and decreasing metabolism and energy expenditure. It is one of the most potent and long- lasting of appetite stimulators. Sample Type: Urine, cell culture supernates, serum, plasma (heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human AGRP/agouti-related. Sensitivity: <4pg/ml Range: 7.8pg/ml-500pg/ml Specificity: Recognizes natural and recombinant Human AGRP/agouti-related. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human AGRP/agouti-related BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for AGRP/agouti-related has been precoated onto 96-well plates. Standards(Expression system for standard: sf21, Immunogen sequence: A21-T132) and test samples are added to the wells, a biotinylated detection specific for AGRP/agouti-related is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human AGRP/agouti-related amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human AGRP antibody, 1x plate Lyophilized recombinant Human AGRP standard, 2x10ng/tube Biotinylated anti-Human AGRP antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
