Human visfatin ELISA Kit

Instructions
Code CSB-E08940h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

CUSABIO's Human visfatin ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the quantitative determination of visfatin levels in serum, plasma, urine, and tissue homogenates. This assay exclusively recognizes human visfatin. The quantitative sandwich ELISA technique of this kit is based on visfatin antibody-visfatin antigen interactions and an HRP colorimetric detection system to detect the levels of visfatin in samples. The intensity of the color is positively proportional to the amount of bound visfatin in the initial step.

This human visfatin ELISA kit has been quality validated for criteria, including sensitivity, specificity, precision, linearity, recovery, and lot-to-lot consistency. Click the product instructions for more details.

Visfatin, also known as nicotinamide phosphoribosyltransferase (NAMPT), is a multifaceted adipokine mainly secreted by the visceral adipose tissue. As a NAD biosynthetic enzyme, NAMPT catalyzes the transformation of nicotinamide into nicotinamide mononucleotide (NMN), which is then converted into NAD+ by nicotinamide/nicotinic acid mononucleotide adenylyltransferase (Nmnat). In addition to its NAD+ biosynthetic activity, NAMPT also acts as a pro-inflammatory cytokine and plays a crucial role in chronic diseases. It also exerts insulin-mimetic effects. Previous studies have shown that NAMPT is effective in activating the insulin signaling cascade, increasing glucose uptake, and inhibiting glucose release. NAMPT is related to glucose metabolism and the pathogenesis of insulin resistance and type II diabetes. NAMPT also causes in vitro endothelial dysfunction and vascular inflammation.

Target Name nicotinamide phosphoribosyltransferase
Alternative Names 1110035O14Rik ELISA Kit; AI314458 ELISA Kit; AI480535 ELISA Kit; DKFZP666B131 ELISA Kit; EC 2.4.2.12 ELISA Kit; MGC117256 ELISA Kit; NAmPRTase ELISA Kit; Nampt ELISA Kit; NAMPT_HUMAN ELISA Kit; Nicotinamide phosphoribosyltransferase ELISA Kit; PBEF 1 ELISA Kit; PBEF ELISA Kit; PBEF1 ELISA Kit; Pre B cell colony enhancing factor 1 ELISA Kit; Pre B cell colony enhancing factor ELISA Kit; Pre B cell enhancing factor ELISA Kit; Pre-B cell-enhancing factor ELISA Kit; Pre-B-cell colony-enhancing factor 1 ELISA Kit; VF ELISA Kit; Visfatin ELISA Kit
Abbreviation NAMPT
Uniprot No. P43490
Species Homo sapiens (Human)
Sample Types serum, plasma, urine, tissue homogenates
Detection Range 0.625 ng/mL-40 ng/mL
Sensitivity 0.156 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Metabolism
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human visfatin in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 87  
Range % 82-93  
1:2 Average % 92  
Range % 87-98  
1:4 Average % 98  
Range % 93-102  
1:8 Average % 102  
Range % 98-106  
Recovery
The recovery of human visfatin spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 100 95-104  
EDTA plasma (n=4) 92 88-96  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
40 1.977 2.050 2.014 1.921  
20 1.526 1.584 1.555 1.462  
10 1.087 1.101 1.094 1.001  
5 0.658 0.703 0.681 0.588  
2.5 0.451 0.421 0.436 0.343  
1.25 0.293 0.301 0.297 0.204  
0.625 0.165 0.159 0.162 0.069  
0 0.096 0.089 0.093    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human NAMPT antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled NAMPT antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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 Q&A
Q:

Please confirm whether this kit can be used with saliva samples and also suggest suitable protocol for saliva sample preparation.

A:
Thanks for your inquiry.
CSB-E08940h ,we used it to test human saliva and the value was ND--2.5ng/ml.
Here is the processing method on saliva sample preparation.You can take it as reference:Remove particulates by centrifugation for 10 minutes at 4000x g at 2-8°C and assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze-thaw cycles. Recommend to use fresh saliva samples.

Target Background

Function
(From Uniprot)
Catalyzes the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, an intermediate in the biosynthesis of NAD. It is the rate limiting component in the mammalian NAD biosynthesis pathway. The secreted form behaves both as a cytokine with immunomodulating properties and an adipokine with anti-diabetic properties, it has no enzymatic activity, partly because of lack of activation by ATP, which has a low level in extracellular space and plasma. Plays a role in the modulation of circadian clock function. NAMPT-dependent oscillatory production of NAD regulates oscillation of clock target gene expression by releasing the core clock component
Gene References into Functions
  1. chemerin and leptin are elevated and omentin-1 and visfatin levels are decreased in Gestational diabetes mellitus women complicated by obesity. PMID: 29337272
  2. fat mass percentage together with HOMA-IR are the most significant predictors of visfatin/NAMPT serum elevation in hyperthyroid patients. PMID: 30184561
  3. Serum levels of visfatin were significant higher in HCC patients than healthy subjects. It was significantly associated with the histology and metastasis. Visfatin induced miR-21 expression and cell migration in HepG2 cells. PMID: 29808800
  4. Knockdown of Nampt expression increased the expression of Aggrecan and Collagen II inhibited by IL-1beta and the autophagic markers, LC3 and Beclin-1, confirmed APO866 induced NP cell autophagy and blocks IL-1beta induction considerably. PMID: 30261504
  5. The study showed for the first time serum omentin and visfatin levels to be decreased after lung transplantation in end-stage lung disease patients PMID: 29277835
  6. We conclude that the low placental visfatin (NAMPT) expression and poor metabolic control in the 3(rd) trimester of pregnancy may have a role in stimulating fetal overgrowth in T1DM pregnancy. PMID: 29769421
  7. no significant differences in genotype frequencies between preterm birth cases and controls; but maternal blood level higher in A allele carriers of rs61330082 PMID: 28482725
  8. higher visfatin levels in erosive hand osteoarthritis (HOA) in comparison to nnonerosive HOA PMID: 29105498
  9. obese subjects with GG genotype of visfatin who had high PUFA-intake diets had lower hip z scores and T scores, unlike the other genotypes PMID: 29355093
  10. Review/Meta-analysis: circulating visfatin levels were significantly higher in patients with rheumatoid arthritis and a positive correlation was suggested with disease activity. PMID: 28205390
  11. Observed NAMPT overexpression in patients with Graves' disease with and without (GO) orbitopathy and its relatively low levels in thyroids of patients with Graves' disease (GD)without eye changes do not confirm causal relationship between NAMPT level and orbitopathy, but this needs further investigation. PMID: 29546048
  12. Visfatin seems to be a good marker of endometrial cancer progress. High visfatin serum level predicts poor prognosis in endometrial cancer patients. PMID: 29516012
  13. NAMPT expression downregulation by mir-206 reduces cell survival of breast cancer cells PMID: 29886033
  14. No correlation between IDH1/2 mutation status and sensitivity for NAMPT inhibitors was observed. Strikingly, higher methylation of the NAPRT promoter was observed in high-grade versus low-grade chondrosarcomas. In conclusion, this study identified NAMPT as a potential target for treatment of chondrosarcoma PMID: 28860121
  15. NAMPT as a key player in BRAFi resistance and melanoma cell plasticity PMID: 29567766
  16. This study has shown Resistin, and Visfatin synergistically increased gastric cancer cell proliferation and enhanced the telomerase gene expression, showing that these two hormones in gastric cancer tissue could cooperatively accelerate cancer cell growth PMID: 29228527
  17. Serum Nampt was increased in diabetic nephropathy patients. PMID: 28699485
  18. NAMPT is highly upregulated in human gingival fibroblasts, while its enzymatic activity acts as a crucial mediator of periodontal inflammation and alveolar bone destruction via regulation of catabolic factors COX-2, MMP1, and MMP3 levels. PMID: 28819322
  19. Results show that adiponectin, resistin and visfatin are present in osteophytes but have no direct influence on osteophyte development. Although these adipokines induce several inflammatory mediators, they do not affect osteoblast- or chondrocyte-related mechanisms of osteophyte development. PMID: 27884778
  20. Cells were treated with 10 nM FK866, an NAMPT inhibitor, resulting in a decrease in the cellular NAD level. PMID: 28714034
  21. data indicate that visfatin induces endothelial progenitor cells apoptosis by increasing the expression of pro-inflammatory mediators partly through the regulation of NF-kappaB. PMID: 28677720
  22. study thus demonstrates that the inhibition of PBEF attenuates oxidative stress and inflammation induced by hyperoxia in EA.hy926 cells. PMID: 28677752
  23. hsa_circ_0005105 can promote extracellular matrix (ECM) degradation by regulating the expression of miR-26a target NAMPT. PMID: 28276108
  24. Careful characterization of the kinetics of NAMPT inhibition in vivo allowed us to optimize dosing to produce sufficient NAD(+) depletion over time that resulted in efficacy in an HCT116 xenograft model. Our data demonstrate that direct phosphoribosylation of competitive inhibitors by the NAMPT enzyme is not required for potent in vitro cellular activity or in vivo antitumor efficacy. PMID: 28468779
  25. Results suggested that visfatin -1535C>T polymorphisms might be associated with decreased risk of HBV-HCC among the ethnic Zhuang population in Guangxi, China. PMID: 27792999
  26. the present study elucidated the pathophysiological role of visfatin in breast cancer and revealed a proliferative and protective role for visfatin PMID: 28442350
  27. serum level associated with increased odds of severe hypertension PMID: 26869250
  28. In this cohort of resistant hypertension patients diagnosed with ambulatory blood pressure measurements, the investigators found an independent correlation between higher visfatin levels and the presence of resistant hypertension and left ventricular hypertrophy. PMID: 28240682
  29. A potential oncogenic role of Nampt in Waldenstrom macroglobulinemia cells. PMID: 27287071
  30. factor with strong correlation with high visfatin values (p=0.001). High Visfatin levels seem to be strongly correlated with the presence and severity of peripheral arterial occlusive disease. PMID: 28073026
  31. These results suggested that the plasma visfatin level is associated with the presence of hepatocellular carcinoma (HCC), and that a higher plasma visfatin level may be important in the pathogenesis of HCC. PMID: 27770715
  32. Increased expression of visfatin are associated with a more aggressive phenotype of colorectal carcinoma patients. It can trigger the epithelial mesenchymal transformation of CRC cells via Akt/GSK-3beta/beta-catenin signals. PMID: 27058759
  33. Results showed that rs34861192 of RETN and rs13237989 of NAMPT synergistically affected the levels of insulin and glycemic index. PMID: 29101068
  34. In summary, NAMPT is produced in human spermatozoa in a maturation-dependent manner. PMID: 27566659
  35. Non-surgical periodontal treatment resulted in a significant reduction in gingival crevicular fluid and serum visfatin levels. Furthermore, visfatin levels increased with inflammation and decreased following periodontal treatment. Our findings suggest that visfatin is an inflammatory biomarker of periodontal disease PMID: 28025432
  36. Accordingly, our results demonstrated that NAMPT is a prognostic marker in melanoma, and the identificationofNAMPT-E2F2-SIRT1 pathway establishes another link between NAMPT and apoptosis events in melanoma, with therapeutic implications for this deadly cancer. PMID: 28919418
  37. systemic circulating NAMPT levels were closely associated with NAMPT release from corresponding cultured PBMCs. In conclusion, NAMPT is selectively increased in states of acute but not chronic inflammation in children PMID: 28837586
  38. It focusing on NAMPT immunometabolic function as a metabokine, NAMPT secretion, NAMPT mechanism of action and possible roles in the cancer micro-environment. PMID: 27128025
  39. Authors show the gene encoding nicotinic acid phosphoribosyltransferase (NAPRT), a second NAD(+)-producing enzyme, is amplified and overexpressed in a subset of common types of cancer, including ovarian cancer. PMID: 28507103
  40. This study shows that expression of SDF-1alpha, eNOS and VEGF were significantly higher in uterine leiomyoma than myometrium with a different expression pattern according to the size of uterine leiomyoma. However, expressions of visfatin and leptin had no significant differences between the two groups. PMID: 28010141
  41. Results indicate that increased serum visfatin levels are associated with poor prognosis of hepatocellular carcinoma (HCC), and visfatin may be a potential therapeutic target of HCC. PMID: 28178643
  42. Nampt expression is a reliable marker of progression in cervical dysplasia and squamous cell carcinoma. PMID: 28870901
  43. Visfatin increases the nuclear translocation of Akt and its binding with ABCC1. These data showed that visfatin can decrease doxorubicin sensitivity of Non-small-cell lung cancer cells via activation of Akt/MRP1. PMID: 28762597
  44. Results suggest a mechanism by which tumor cells protect themselves against glucose deprivation-induced oxidative stress by utilizing nicotinamide phosphoribosyltransferase (NAMPT) to maintain NADPH levels. PMID: 26568303
  45. The combination of upregulation of the NAD(+) de novo synthesis pathway through QPRT over-expression and NAMPT mutation confers resistance to GMX1778, but the cells are only partially resistant to next-generation NAMPT inhibitors. The resistance mechanisms uncovered herein provide a potential avenue to continue exploration of next generation NAMPT inhibitors to treat neoplasms in the clinic. PMID: 28756225
  46. this study shows that upstream visfatin SNPs could potentially affect phenotypic outcome in obese children through alteration of circulating visfatin level. PMID: 27887573
  47. Lack of changes in serum visfatin level despite the changes in visfatin mRNA expression of adipose tissue suggests paracrine effect of visfatin rather than endocrine PMID: 26486101
  48. Ascending aorta from aortic aneurysm patient,s revealed an inverse relationship between smooth muscle cell NAMPT content and aortic diameter. PMID: 28356339
  49. Visfatin significantly correlated with BMI and hot flashes number in postmenopausal women with metabolic syndrome. PMID: 27126198
  50. Single nucleotide polymorphism in the NAMPT gene is associated with inflammation in chronic kidney disease. PMID: 27190335

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Subcellular Location Nucleus, Cytoplasm, Secreted
Protein Families NAPRTase family
Tissue Specificity Expressed in large amounts in bone marrow, liver tissue, and muscle. Also present in heart, placenta, lung, and kidney tissues.
Database Links

HGNC: 30092

OMIM: 608764

KEGG: hsa:10135

STRING: 9606.ENSP00000222553

UniGene: Hs.489615

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