Vitamin B3 (VB3) ELISA Kit

Sample

Serum, plasma and other biological fluids

Detection Method

cELISA

Intended Use

The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of VB3 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.

Contents of Kit

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

Storage

1. For unused kit: The whole kit could be stored at -20°C in shelf life, while up to one month at 4°C. For experiment convenience, reagents could also be stored seperately, Standard, Detection Reagent A, Detection Reagent B and 96-well strip plate should be stored at -20°C while the others could be at 4°C.
2. For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.
Note:
It is highly recommended to use the remaining reagents within 1 month provided this is prior to the expiration dateof the kit. For the expiration date of the kit, please refer to the label on the kit box. All components are stable up tothe expiration date.

Performance Characteristics

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

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level VB3 were tested 20times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level VB3 were tested on3 different plates, 8 replicates in each plate. CV(%) = SD/mean×100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Detection Range

246.9-20,000pg/mL. The standard curve concentrations used for the ELISA's were 20,000pg/mL, 6,666.7pg/mL,2,222.2pg/mL, 740.7pg/mL, 246.9pg/mL.

Sensitivity

The minimum detectable dose of VB3 is typically less than 88.3pg/mL.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentrationthat could be differentiated from zero. It was determined by subtracting two standard deviations to the meanoptical density value of twenty zero standard replicates and calculating the corresponding concentration.

Standard Curve

In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the log ofconcentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is thedependent variable. The O.D. values of the standard curve may vary according to the conditions of assayperformance (e.g. operator, pipetting technique, washing technique or temperature effects). Typical standardcurve below is provided for reference only.