25-HVD3(25-Hydroxy Vitamin D3) ELISA Kit (E-EL-0015)
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For research use only.
Product Summary
| Sensitivity | 1.88 ng/mL |
| Detection Range | 3.13-200 ng/mL |
| Sample Volume | 50 μL |
| Total Assay Time | 2 h 30 min |
| Reactivity | Universal |
| Specificity | This kit recognizes Universal 25-HVD3 in samples.No significant cross-reactivity or interference between Universal 25-HVD3 and analogues was observed |
| Recovery | 80%-120% |
| Sample Type | Serum, plasma and other biological fluids |
| Detection Method | Colorimetric method, ELISA, Competitive |
| Assay Type | Competitive-ELISA |
| Size | 96T / 48T / 24T / 96T*5 / 96T*10 |
| Storage | 2-8℃ |
| Expiration Date | 12 months |
Performance
| Typical data |
The following data was generated by the Quality Control Department, under controlled laboratory conditions (ambient temperature: 18-25 °C, relative humidity: 35-75%) using standardized procedures (TMB reaction at 37 °C in the dark for 15 minutes, followed by termination and OD measurement). These values are provided for reference only.
Actual results may vary due to differences in laboratory conditions, operator technique, and equipment. Users are required to generate a standard curve using their own experimental data.
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| Precision |
Intra-assay Precision (Within-run Precision): Three samples representing low, mid, and high concentrations of Human IL-6 were tested 20 times on a single plate.
Inter-assay Precision (Between-run Precision): Three samples representing low, mid, and high concentrations of Human IL-6 were tested on three separate plates, with 20 replicates per plate, to assess variability among assays.
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| Recovery |
The recovery of Universal 25-HVD3 was evaluated by spiking samples at low, mid, and high concentrations across the assay range in various sample matrices.
The assay performance was assessed by comparing the measured concentrations to the expected spiked amounts to determine the percent recovery.
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| Linearity |
Linearity of the assay was evaluated by spiking samples with high concentrations of Universal 25-HVD3 and performing serial dilutions using Standard & Sample Diluent to produce concentrations spanning the assay's dynamic range.
The measured values were then compared to the expected concentrations to assess the linearity of response.
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| Stability |
Each kit batch is subjected to accelerated stability testing and real-time stability monitoring. Sample performance is evaluated after storage at 37 °C for 10 days to assess the impact of elevated temperature on assay reliability and reagent integrity.
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Test Principle
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal 25-HVD3. During the reaction, Universal 25-HVD3 in the sample or standard competes with a fixed amount of Universal 25-HVD3 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal 25-HVD3. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Universal 25-HVD3 in tested samples can be calculated by comparing the OD of the samples to the standard curve.
Background
25-hydroxyvitamin D, also known as 25-hydroxyvitamin D3, is the main form of vitamin D in the body. It is produced by cholecalciferol (vitamin D3) under the induction of 25-hydroxylase in the liver.The main functions of 25-hydroxyvitamin D are to promote the absorption of calcium and phosphorus by the small intestinal mucosa, promote osteoclast activity and enhance the mobilization of bone calcium by the parathyroid glands, and promote the reabsorption of calcium by the renal tubules, etc.
| Research Area | Metabolism , Developmental Biology |
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