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 Nampt (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) (mouse/rat) 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 mouse and rat Nampt. It does not cross-react with human Nampt, human adiponectin, mouse adiponectin, mouse resistin, mouse vaspin, mouse RBP4, mouse GPX3, mouse progranulin, mouse IL-33, mouse clusterin, mouse ANGPTL3, mouse ANGPTL4, mouse ANGPTL6, mouse leptin, mouse TNF-a. The assay range is 0.5-32ng Nampt/ml and a detection limit of 50pg/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 mouse and rat Nampt.Sensitivity:50pg/mlRange:0.5-32ng Nampt/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): 1x150ulMouse Nampt Standard (lyophilized, 64ng): 1 vialMouse 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.