Agouti-related Protein (AGRP) BioAssay(TM) ELISA Kit (Mouse)

This gene encodes an antagonist of the melanocortin-3 and melanocortin-4 receptor. It appears to regulate hypothalamic control of feeding behavior via melanocortin receptor and/or intracellular calcium regulation, and thus plays a role in weight homeostasis. Mutations in this gene have been associated with late on-set obesity. Agouti-related Protein (AGRP) BioAssay(TM) ELISA Kit a sandwich ELISA intended for the quantitative determination of AGRP in mouse serum, plasma and tissue homogenates. , Detection Range:, 15.6-1000pg/ml, , Intra-assay Precision: %CV <8 , Inter-assay Precision: %CV <10, Sensitivity: The minimum detectable dose of mouse AGRP is typically less than 3.9pg/ml., The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined the mean O.D value of 20 replicates of the zero standard added by their three standard deviations. Specificity: This assay has high sensitivity and excellent specificity for detection of mouse AGRP. No significant crossreactivity or interference between mouse AGRP and analogs was observed., Note: Limited by current skills and knowledge, it is impossible for us to complete the crossreactivity detection between mouose AGRP and all the analogs. Therefore, crossreaction may still exist. Kit Components: 351588A: Microtiter Strips, 1x48 wells or 1x96 wells. Pre-coated, ready to use. , 351588B: Standard, 1x1vial or 2x1vial, 351588C: AGRP (Biotin), 100X, 1x60ul or 1x120ul, 351588D: Avidin (HRP), 100X, 1x60ul or 1x120ul, 351588E: Biotin Diluent, 1x7.5ml or 1x15ml, 351588F: Avidin (HRP) Diluent, 1x7.5ml or 1x15ml, 351588G: Sample Diluent, 1x25ml or 1x50ml, 351588H: Wash Buffer, 25X, 1x10ml or 1x20ml, 351588J: TMB Substrate, 1x5ml or 1x10ml, 351588K: Stop Solution, 1x5ml or 1x10ml, Note: The Stop Solution (351588K) provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store kit at 4°C. The unopended kit is stable for 6 months after receipt. Once opened, the kit is stable up to 4 weeks at 4°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Prepare reagents, standards and samples as instructed. 2. Add 100ul standard or sample to each well. Incubate for 2 hours at 37°C, 3. Remove the liquid of each well do not wash., 4. Add 100ul AGRP (Biotin), 1X to each well. Incubate for 1 hour at 37°C, 5. Aspirate and wash 3 times., 6. Add 100ul Avidin (HRP), 1X to each well. Incubate for 1 hour at 37°C, 7. Aspirate and wash 5 times., 8. Add 90ul TMB substrate to each well Incubate for 15-30 minutes at 37°C. Protect from light, 9. Add 50ul Stop Solution to each well. Read at 450nm within 5 minutes.