Nampt (mouse) (rec.) (enzymatically active)
Method: The protein Nampt (mouse) (rec.) (1µM) is tested for activity in vitro using the Cyclex NAMPT Colorimetric Assay Kit (Cyclex / MBL, CY-1251) in the presence of different concentrations of CHS-828 (dissolved in DMSO). The IC50 of CHS-828 is 40nM in this experiment.
Method: The protein Nampt (mouse) (rec.) is tested for activity in vitro using the Cyclex NAMPT Colorimetric Assay Kit (Cyclex; MBL, CY-1251). Nampt is a rate-limiting enzyme in the salvage pathway of NAD+ biosynthesis. This Nampt assay measures the formation of nicotinamide > nicotinamide mononucleotide (NMN) > nicotinamide adenine dinucleotide (NAD) > NADH. The newly formed NADH is then oxidized resulting in the reduction of a tetrazolium salt substrate (WST-1) to a highly-colored formazan which absorbs at 450nm (see manufacturer‘s protocol). The activity of the Nampt (mouse) (rec.) (enzymatically active) (AG-40B-0179) (100nM) is shown at different times. As negative control, an unrelated protein (100nM) is used.
|Synonyms||Visfatin; PBEF; Pre-B Cell Colony Enhancing Factor 1; PBEF1; Nicotinamide Phosphoribosyltransferase; NAmPRTase|
|Source/Host||HEK 293 cells|
Mouse Nampt (visfatin/PBEF) (aa 1-491) is fused at the N-terminus to a FLAG®-tag.
Enzymatically active to convert nicotinamide + phosphoribosyltransferase (PRPP) to nicotinamide mononucleotide (NMN).
0.5µg to 5µg of Nampt (mouse) (rec.) (enzymatically active) show activity in a colorimetric Nampt assay.
|MW||~55kDa (SDS-PAGE); ~100-110 kDa (Dimer) (SEC)|
|Endotoxin Content||<0.01EU/μg purified protein (LAL test; Lonza).|
|Formulation||Liquid. Contains PBS.|
|Other Product Data||
UniProt link Q99KQ4: Nicotinamide phosphoribosyltransferase (mouse)
FLAG is a registered trademark of Sigma-Aldrich Co.
|Shipping and Handling|
|Short Term Storage||-20°C|
|Long Term Storage||-80°C|
After opening, prepare aliquots and store at -80°C.
Avoid freeze/thaw cycles.
Stable for at least 6 months after receipt when stored at -80°C.
Working aliquots are stable for up to 3 months when stored at -20°C.
|Product Specification Sheet|
Nicotinamide phosphoribosyltransferase (NAmPRTase or Nampt) also known as pre-B cell colony-enhancing factor 1 (PBEF1) or visfatin is the rate-limiting enzyme in the nicotinamide adenine dinucleotide (NAD+) salvage pathway that converts nicotinamide to nicotinamide mononucleotide in mammals to enable NAD+ biosynthesis. In addition it has been shown to be an adipokine that is localized in the bloodstream and has various functions, including the promotion of vascular smooth muscle cell maturation and inhibition of neutrophil apoptosis. It activates insulin receptor and has insulin-mimetic effects, lowering blood glucose and improving insulin sensitivity. The protein is highly expressed in visceral fat and serum levels of the protein correlate with obesity. The recombinant Nampt (mouse) (rec.) (enzymatically active) (Prod. No. AG-40B-0179) shows activity to convert nicotinamide + phosphoribosyltransferase (PRPP) to nicotinamide mononucleotide (NMN). It migrates as a homodimer on a size exclusion chromatography (SEC) column.