CLIA Kit for Vitamin D2 (VD2)

The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to Vitamin D2 (VD2) . A competitive inhibition reaction is launched between biotin labeled Vitamin D2 (VD2) and unlabeled Vitamin D2 (VD2) (Standards or samples) with the pre-coated antibody specific to Vitamin D2 (VD2) . After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Vitamin D2 (VD2) in the sample. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is reverse proportional to the Vitamin D2 (VD2) level in the sample or standard.

1. Prepare all reagents, samples and standards; <br/> 2. Add 50&micro; L standard or sample to each well.<br/> &nbsp; &nbsp; &nbsp; &nbsp; And then add 50&micro; L prepared Detection Reagent A immediately. <br/> &nbsp; &nbsp; &nbsp; &nbsp; Shake and mix. Incubate 1 hour at 37&deg; C; <br/> 3. Aspirate and wash 3 times; <br/> 4. Add 100&micro; L prepared Detection Reagent B. Incubate 30 minutes at 37&deg; C; <br/> 5. Aspirate and wash 5 times; <br/> 6. Add 100&micro; L Substrate Solution. Incubate 10 minutes at 37&deg; C; <br/> 7. Read RLU value immediately.<br/>

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Vitamin D2 (VD2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Vitamin D2 (VD2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV Inter-Assay: CV Inter-Assay: CV

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.

This assay has high sensitivity and excellent specificity for detection of Vitamin D2 (VD2) . No significant cross-reactivity or interference between Vitamin D2 (VD2) and analogues was observed.