ELISA Kit for Visfatin (VF)

Nampt; NAmPRTase; PBEF; PBEF1; Pre-B-Cell Colony Enhancing Factor; Nicotinamide Phosphoribosyltransferase

Specificity

This assay has high sensitivity and excellent specificity for detection of Visfatin (VF).
No significant cross-reactivity or interference between Visfatin (VF) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Visfatin (VF) and the recovery rates were calculated by comparing the measured value to the expected amount of Visfatin (VF) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-101 94
EDTA plasma(n=5) 94-102 99
heparin plasma(n=5) 82-93 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Visfatin (VF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Visfatin (VF) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Visfatin (VF) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 91-101% 90-101% 86-98% 88-104%
EDTA plasma(n=5) 96-104% 89-97% 82-93% 78-92%
heparin plasma(n=5) 91-99% 79-97% 81-93% 81-92%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Brazilian Journal of Biomoricity EFFECT OF 8 WEEKS ENDURANCE TRAINING ON PLASMA VISFATIN IN MIDDLE-AGED MEN Brjb: source
Diabetes Research and Clinical Practice The relationship between visfatin and HOMA-IR in hypertensive patients, and the effect of antihypertensive drugs on visfatin and HOMA-IR in hypertensive patients with insulin resistance ScienceDirect: S0168822711002956
J Physiol Pharmacol. Early-effect of bariatric surgery (Scopinaro method) on intestinal hormones and adipokines in insulin resistant Wistar rat. Pubmed: 24304571
Endocr J. CTRP3 modulates the expression and secretion of adipokines in 3T3-L1 adipocytes Pubmed:25168658
Exp Ther Med. Plasma visfatin levels and mRNA expression of visfatin in peripheral blood mononuclear cells and peripheral blood monocyte-derived macrophages from normal weight females with polycystic ovary syndrome Pubmed:Pmc3991522
Eur J Cancer Prev. Visfatin derived from ascites promotes ovarian cancer cell migration through Rho/ROCK signaling-mediated actin polymerization. Pubmed:25055182
Genome Visfatin −948G/T and resistin −420C/Gpolymorphisms in Egyptian type 2 diabetic patients with and without cardiovascular diseases Nrcresearchpress:Source
PLOS ONE Antidiabetic Effect of Galantamine: Novel Effect for a Known Centrally Acting Drug PubMed: 26262991
Alcoholism and Drug Addiction Changes in concentration of visfatin during four weeks of inpatient treatment of alcohol dependent males Science: Article
Cytokine Variation in blood levels of hormones in obese patients following weight reduction induced by endoscopic and surgical bariatric therapies PubMed: 26539806
Genome Genetic Variations in Visfatin and Resistin Genes in Egyptian Type 2 Diabetic Patients With and Without Complications PubMed: 25120107
Acta Histochemica Favorable outcomes of hydroxychloroquine in insulin resistance may be accomplished by adjustment of the endothelial dysfunction as well as the skewed balance of adipokines science:S0065128116301234
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