Nampt, nicotinamide phosphoribosy-ltransferase, is the rate limiting enzyme of the mammalian NAD biosynthesis pathway. The circulating NMN (nicotinamide mononucleotide) and NAD are taken up by beta cells and converted to NAD by Nmnat (nicotinamide mononucleotide adenyltransferase) and intracellular Nampt. The Nampt plays a critical role in enhancing life span and protecting against oxidative damage. The Nampt (Visfatin/PBEF) (human) Intracelluar ELISA Kit is to be used for the in vitro quantitative determination of intracellular mouse or rat Nampt (Visfatin/PBEF). This assay is a sandwich ELISA which utilizies a 96-well microtiter plate which was pre-coated with a monoclonal antibody and a purified polyclonal detection antibody. A HRP-conjugated anti-IgG and TMB (3,3',5,5'-tetramethylbenzidine) is added to generate a color intensity directly proportional to the concentration of Nampt in the samples. This ELISA is specific for the measurement of natural and recombinant human Nampt. It does not cross-react with human adiponectin, human resistin, human RELM-b, human leptin, human GPX3, human ANGPTL4, human FABP4, human ANGPTL6, human PAI1. The assay range is 0.063-16ng Nampt/ml and a detection limit of 30pg/ml (based on adding two standard deviations to the mean value of the zero standard).Sample Type:Cell lysatesIntended Use:For the measurement of natural and recombinant human Nampt.Sensitivity:30pg/mlRange:0.063-16ng Visfatin/mlSpecificity:MammalianTest Principle:This assay is a sandwich ELISA which utilizies a 96-well microtiter plate which was pre-coated with a monoclonal antibody and a purified polyclonal detection antibody. A HRP-conjugated anti-IgG and TMB (3,3',5,5'-tetramethylbenzidine) is added to generate a color intensity directly proportional to the concentration of Nampt in the samples.Kit Components:Pre-coated Microtiter Plate: 12x8 well stripsWash Buffer (10X): 1x50mlDiluent (5X):1x50mlDetection Antibody: 1x12mlDetector 100X (Hrp conjugated anti-IgG): 1x150ulHuman Nampt Standard (lyophilized, 32ng): 1 vialHuman Nampt QC Sample (lyophilized): 1 vialTMB Substrate Solution: 1x12mlStop Solution: 1x12mlPlate Sealers: 3 eachLysis Buffer (10X): 1x12mlStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.