ELISA Kit for Vanin 1 (VNN1) Rattus norvegicus (Rat) Sandwich ELISA

Pantetheinase; Pantetheine hydrolase; Tiff66; Vascular non-inflammatory molecule 1

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  • ELISA Kit for Vanin 1 (VNN1) Packages (Simulation)
  • ELISA Kit for Vanin 1 (VNN1) Packages (Simulation)
  • ELISA Kit for Vanin 1 (VNN1) Results demonstration
  • SEC598Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Vanin 1 (VNN1) and the recovery rates were calculated by comparing the measured value to the expected amount of Vanin 1 (VNN1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-104 96
EDTA plasma(n=5) 91-103 94
heparin plasma(n=5) 89-98 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vanin 1 (VNN1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vanin 1 (VNN1) 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 Vanin 1 (VNN1) 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) 98-105% 88-96% 80-104% 80-93%
EDTA plasma(n=5) 86-103% 82-94% 93-105% 78-101%
heparin plasma(n=5) 85-105% 84-98% 88-102% 84-95%

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 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Vanin 1 (VNN1)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Vanin 1 (VNN1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Vanin 1 (VNN1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Vanin 1 (VNN1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Vanin 1 (VNN1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Vanin-1; a potential biomarker for nephrotoxicant-induced renal injuryScienceDirect: S0300483X11003234
  • Urinary vanin-1 as a novel biomarker for early detection of drug-induced acute kidney injuryAspet: 192807
  • Urinary vanin-1 as a novel biomarker for early detection of drug-induced acute kidney injury.PubMed: 22399813
  • Early detection of renal injury using urinary vanin-1 in rats with experimental colitisPubMed: 23307618
  • Effects of Exercise and Ferula Gummosa on Apelin of Cardiac and Kidney Tissues in L-NAME induced Hypertension in RatsJm.um: Source
  • Value of vanin-1 assessment in adult patients with primary immune thrombocytopeniaPubmed: 23534352
  • Early detection of renal injury using urinary vanin-1 in rats with experimental colitis.Pubmed: 23307618
  • Urinary Kim-1 is a sensitive biomarker for the early stage of diabetic nephropathy in Otsuka Long-Evans Tokushima Fatty ratsSagepub:Source
  • The effect of activated charcoal on adenine-induced chronic renal failure in ratsPubmed:24412558
  • Early urinary biomarkers for renal tubular damage in spontaneously hypertensive rats on a high salt intakePubMed: 26376947
  • Citrate Attenuates Adenine-Induced Chronic Renal Failure in Rats by Modulating the Th17/Treg Cell BalancePubMed: 26253296
  • Early prediction of cisplatin-induced nephrotoxicity by urinary vanin-1 in patients with urothelial carcinomaPubmed:27317936
  • Nanosized titanium dioxide resulted in the activation of TGF-β/Smads/p38MAPK pathway in renal inflammation and fibration of micePubmed:26850371
  • Citrate attenuates vascular calcification in chronic renal failure rats.pubmed:28332248
  • Early urinary biomarkers of renal tubular damage by a high-salt intake independent of blood pressure in normotensive rats.pubmed:29027259
  • Gum Acacia Improves Renal Function and Ameliorates Systemic Inflammation, Oxidative and Nitrosative Stress in Streptozotocin-Induced Diabetes in Rats …Pubmed:29550811
  • Vanin-1 in Renal Pelvic Urine Reflects Kidney Injury in a Rat Model of HydronephrosisPubmed: 30332759
  • Involvement of Vanin-1 in Ameliorating Effect of Oxidative Renal Tubular Injury in Dahl-Salt Sensitive RatsPubmed: 31514290
  • Urinary vanin‐1 associated with chronic kidney disease in hypertensive patients: A pilot studyPubmed: 32869948
  • Association of urinary vanin©\1 with kidney function decline in hypertensive patients34029440
  • Plasma vanin-1 as a novel biomarker of sepsis for trauma patients: a prospective multicenter cohort study33624223

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