NAMPT ELISA Kit

INTENDED USE

Human NAMPT ELISA kit can measure concentrations of nicotinamide phosphoribosyltransferase (NAMPT, Visfatin, NMPRTase, PBEF1, Pre-B cell-enhancing factor) present is plasma, serum, biological fluids or tissue homogenate samples. This assay has a sensitivity detection limit of 0.188 ng/ml.

INTRODUCTION

Nicotinamide phosphoribosyltransferase (NAMPT) is a protein enzyme that plays a role, in the production of nicotinamide adenine dinucleotide (NAD+) an essential coenzyme is found in all living cells. The enzyme is responsible for a step in generating NAD+ from nicotinamide. By regulating NAD+ biosynthesis it has an impact on metabolism, energy production and signaling pathways.

The NAMPT-gene encodes the NAMPT enzyme. It converts nicotinamide into nicotinamide mononucleotide (NMN) which then transforms into NAD+. In NAD+ metabolism NAMPT is believed to be the factor that determines its rate. When there is a demand for NAD+, such as during exercise fasting or DNA damage it gets upregulated. Maintaining levels of NAD+ supports function and ensures energy balance.

Apart from its role NAMPT also acts as a hormone known as visfatin. Visfatin is released by fat and mimics insulins actions by reducing blood glucose levels. The diverse effects of NAMPT/visfatin have implications for health span. Contributes to conditions like obesity, aging, cancer and neurodegenerative disorders. Researchers are exploring the potential of inhibitors targeting NAMPT to deplete NAD+ as a strategy for tumor effects, in cancer therapy.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti- NAMPT antibody.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this NAMPT ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.312, 0.625, 1.25, 2.5, 5, 10, 20 ng/ml.
Reactivity: Human
Sensitivity: 0.188 ng/ml
Range: 0.313 – 20 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for NAMPT, no cross reactivity or interference between NAMPT and analogues was detected.
– Recovery: Serum (87 – 105%), EDTA Plasma (89 – 102%), Heparin Plasma (88 – 103%).
– Linearity: Serum (92 – 104%), EDTA Plasma (85 – 100%), Heparin Plasma (81 – 99%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. NAD⁺ Metabolism Maintains Inducible PD-L1 Expression to Drive Tumor Immune Evasion. Cell Metab. (2021) 33 (1): 110-127. e5. Lv H., et al.
2. Nicotinamide Phosphoribosyltransferase as a Key Molecule of the Aging/Senescence Process. Int J Mol Sci. (2021) 22 (7): 3709. Khaidizar F.D., et al.
3. NAMPT in regulated NAD biosynthesis and its pivotal role in human metabolism. Curr Med Chem. (2011) 18 (13): 1947-61. Burgos E.S.
4. The role of visfatin in pathogenesis of gestational diabetes (GDM). Ginekol Pol. (2018) 89 (9): 518-521. Radzicka S., et al.
5. Boosting NAD⁺ with a small molecule that activates-NAMPT. Nat Commun. (2019) 10 (1): 3241. Gardell S.J., et al.

ADDITIONAL INFORMATION

• Full Name: NAMPT ELISA Kit (Nicotinamide Phosphoribosyltransferase)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 0.188 ng/ml

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