Multiplex Assay Kit for 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array)

This assay has high sensitivity and excellent specificity for detection of 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Matrices listed below were spiked with certain level of 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array) in samples.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 25-Hydroxyvitamin D3 (HVD3) ,etc. by CBA (Cytometric Bead Array) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

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.

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.