Multiplex Assay Kit for Vanin 1 (VNN1) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

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

(Note: Up to 8-plex in one testing reaction)

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  • Multiplex Assay Kit for Vanin 1 (VNN1) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Vanin 1 (VNN1) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Vanin 1 (VNN1) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMC598Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-98 88
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 86-98 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vanin 1 (VNN1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 81-94% 78-89% 85-95% 86-97%
EDTA plasma(n=5) 80-91% 86-99% 80-102% 89-99%
heparin plasma(n=5) 88-102% 80-89% 80-101% 90-98%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:VNN1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Multiplex Assay Kit for Vanin 1 (VNN1) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

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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