Typical Testing Data/Standard Curve (for reference only)
Nampt elisa kit
Nampt (Visfatin/PBEF) (Mouse/Rat) ELISA Kit
Typical Testing Data/Standard Curve (for reference only)
Typical Testing Data/Standard Curve (for reference only)
Principle of the Assay: This assay is a sandwich Enzyme Linked-Immunosorbent Assay (ELISA) for the measurement of human Nampt in cells. A monoclonal antibody specific for Nampt has been pre-coated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, Nampt is recognized by the addition of a purified polyclonal antibody specific for Nampt (Detection Antibody). After removal of excess polyclonal antibody, HRP conjugated anti-rabbit IgG (HRP) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450nm after acidification and is directly proportional to the concentration of Nampt in the samples!!Background/Introduction: Nampt, nicotinamide phosphoribosyl-transferase, is the rate-limiting enzyme of the mammalian NAD, nicotinamide adenine dinucleotide, biosynthesis pathway from nicotinamide to NMN, nicotinamide mononucleotide (1), which is further converted to NAD by nicotinamide/nicotinic acid mononucleotide adenyltransferase abbreviated Nmnat. Nampt was originally identified in Haemophilus ducreyi (2), and was then found to have a significant homology to the mammalian pre-B cell colony-enhancing factor (PBEF) (3) or visfatin (4), which is also called extracellular Nampt (eNampt) (5) due to the fact that it is found as a circulating form in human serum and secreted from differentiated adipocytes. Although visfatin was originally reported as an insulin-mimicking hormone by its capability of binding to and activating the insulin receptor, subsequent studies do not support this observation (5, 6). However, Revollo et al. observed that haplodeficiency and chemical inhibition of Nampt caused defects in NAD biosynthesis and glucose-stimulated insulin secretion in pancreatic islets in vivo and in vitro and plasma visfatin and NMN levels were reduced in Nampt heterozygous females (5). This study proposed a model that NMN exists in high amounts in plasma, presumably derived from nicotinamide with help of eNampt. This circulating NMN and nicotinamide are uptaken by beta cells via unknown transport mechanism (s) are converted to NAD by Nmnat and intracellular Nampt (hereinafter abbreviated iNampt), respectively, concluding that Nampt-mediated systemic NAD biosynthesis is critical for insulin secretion presumably via a NAD-dependent histonedeacetylase, Sirt1. Since it has been shown that NAD(+) levels in mitochondria remain at physiological levels following genotoxic stress and can maintain cell viability even when nuclear and cytoplasmic pools of NAD(+) are depleted (7), the NAD(+) biosynthetic enzyme Nampt plays a critical role in enhancing life span and protecting against oxidative cell damage. Therefore, measurement of iNampt located at different cell compartments like cytoplasm, mitochondria, and nucleus may be able to give us some biological clue on the intracellular functions of Nampt.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a protein that catalyzes the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, one step in the biosynthesis of nicotinamide adenine dinucleotide. The protein belongs to the nicotinic acid phosphoribosyltransferase (NAPRTase) family and is thought to be involved in many important biological processes, including metabolism, stress response and aging. This gene has a pseudogene on chromosome 10. [provided by RefSeq, Feb 2011]
Uniprot Description
PBEF: an ubiquitous protein originally described as a cytokine which acts on early B-lineage precursor cells. Other reports indicate that PBEF is not a cytokine-like secreted protein but an intracellular protein associated with the cell cycle. Contains a catalytically active nicotinate phosphoribosyltransferase domain, an enzyme involved in nicotinamide adenine dinucleotide (NAD) biosynthesis.
Protein type: Cofactor and Vitamin Metabolism - nicotinate and nicotinamide; Transferase; Cell cycle regulation; Cytokine; EC 2.4.2.12
Chromosomal Location of Human Ortholog: 7q22.3
Cellular Component: cell junction; cytosol; extracellular space; nucleoplasm
Molecular Function: cytokine activity; drug binding; nicotinamide phosphoribosyltransferase activity; nicotinate-nucleotide diphosphorylase (carboxylating) activity; protein binding; protein homodimerization activity
Biological Process: cell-cell signaling; circadian regulation of gene expression; circadian rhythm; female pregnancy; insulin receptor signaling pathway; NAD biosynthetic process; NAD metabolic process; nicotinamide metabolic process; positive regulation of cell proliferation; positive regulation of nitric-oxide synthase biosynthetic process; positive regulation of smooth muscle cell proliferation; positive regulation of transcription from RNA polymerase II promoter; response to organic cyclic substance; signal transduction; vitamin metabolic process; water-soluble vitamin metabolic process
Research Articles on Nampt
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Product Notes
The Nampt nampt (Catalog #AAA495239) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Nampt, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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