Products specifications
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Species
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General
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Format
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96T
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Sample Type
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Serum, plasma and other biological fluids.
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Assay Length
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2.5 h
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Detection Range
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2.47-200ng/mL. The standard curve concentrations used for the ELISA’s were 200ng/mL, 66.67ng/mL, 22.22ng/mL, 7.41ng/mL, 2.47ng/mL
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Sensitivity
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0.88ng/mL
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Specificity
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This assay has high sensitivity and excellent specificity for detection of 25-Hydroxyvitamin D3 (HVD3).
No significant cross-reactivity or interference between 25-Hydroxyvitamin D3 (HVD3) and analogues was observed.
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Intra-Assay Precision
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<10%
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Inter-Assay Precision
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<12%
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Storage Temperature
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-20 °C
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Shipping Conditions
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2-8 °C
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Handling Instruction
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The activity loss rate (stability) of this kit is less than 5% within the expiration date under appropriate storage conditions. To minimize extra influence on the stability, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. When not in use, kit components should be refrigerated. All reagents should be brought to room temperature before use. It is recommended that all standards, controls, and samples be run in duplicate. Do not mix or interchange different reagent lots from various kit lots.
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Test Principle
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This assay employs the competitive inhibition enzyme immunoassay technique. An antibody specific for 25-Hydroxyvitamin D3 (HVD3) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between Horseradish Peroxidase (HRP) labeled 25-Hydroxyvitamin D3 (HVD3) and unlabeled 25-Hydroxyvitamin D3 (HVD3) (Standards or samples) with the pre-coated antibody specific for 25-Hydroxyvitamin D3 (HVD3). After incubation the unbound conjugate is washed off. The amount of bound HRP conjugate is reverse proportional to the concentration of 25-Hydroxyvitamin D3 (HVD3) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of 25-Hydroxyvitamin D3 (HVD3) in the sample.
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Notes
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All kits are custom made on-demand. Please allow 10-15 working days for production and 2-3 days for shipment.
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Legal Information
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Marketed under license from USCN Life Science Inc.
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| Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 |
| Standard | 2 |
| Detection Reagent A | 1×120µL |
| Detection Reagent B | 1×120µL |
| TMB Substrate | 1×9mL |
| Wash Buffer (30 × concentrate) | 1×20mL |
| Plate sealer for 96 wells | 4 |
| Standard Diluent | 1×20mL |
| Assay Diluent A | 1×12mL |
| Assay Diluent B | 1×12mL |
| Stop Solution | 1×6mL |
| Instruction manual | 1 |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 25-Hydroxyvitamin D3 (HVD3) 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) | 82-91% | 81-104% | 87-99% | 91-101% |
| EDTA plasma(n=5) | 80-95% | 83-102% | 79-99% | 97-104% |
| heparin plasma(n=5) | 86-104% | 90-97% | 93-105% | 78-98% |
Matrices listed below were spiked with certain level of recombinant 25-Hydroxyvitamin D3 (HVD3) and the recovery rates were calculated by comparing the measured value to the expected amount of 25-Hydroxyvitamin D3 (HVD3) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 99-105 | 102 |
| EDTA plasma(n=5) | 78-89 | 85 |
| heparin plasma(n=5) | 93-105 | 98 |