E01D0002 Human 1,25 dihydroxyvitamin D3 ELISA kit
Human 1,25 dihydroxyvitamin D3 ELISA kit is suitable for detecting samples from the human species. 1,25 dihydroxy vitamin D3 can also be called 1,25(OH)2D3.
Specifications of Human 1,25 dihydroxyvitamin D3 ELISA kit
| Product Information | |
| Cat. No. | E01D0002 |
| Product Name | Human 1,25 dihydroxyvitamin D3 ELISA kit |
| Species | Human |
| Product Size | 48 Tests / 96 Tests |
| Concentration | 25-500 pg/mL |
| Sensitivity | 1.0 pg/mL |
| Principal | Competitive ELISA |
| Sample Volume | 100 ul |
| Sample Type | Serum, plasma, cell culture supernatants, body fluid and tissue homogenate |
| Assay Time | 90 minutes |
| Platform | Microplate Reader |
| Conjugate | HRP |
| Detection Method | Colorimetric |
| Storage | 2-8°C |
| Kit Components | ||
| MATERIALS | SPECIFICATION | QUANTITY |
| MICROTITER PLATE | 96 wells | stripwell |
| ENZYME CONJUGATE | 6.0 mL | 1 vial |
| STANDARD A (0.5mL) | 0 pg/mL | 1 vial |
| STANDARD B (0.5mL) | 25 pg/mL | 1 vial |
| STANDARD C (0.5mL) | 50 pg/mL | 1 vial |
| STANDARD D (0.5mL) | 100 pg/mL | 1 vial |
| STANDARD E (0.5mL) | 250 pg/mL | 1 vial |
| STANDARD F (0.5mL) | 500 pg/mL | 1 vial |
| SUBSTRATE A | 6 mL | 1 vial |
| SUBSTRATE B | 6 mL | 1 vial |
| STOP SOLUTION | 6 mL | 1 vial |
| WASH SOLUTION (100 x) | 10 mL | 1 vial |
| BALANCE SOLUTION | 3 mL | 1 vial |
| Principle of the Assay | |
1,25 (OH)2D3 ELISA kit applies the competitive enzyme immunoassay technique utilizing an anti-1,25 (OH)2D3 antibody and an 1,25 (OH)2D3-HRP conjugate. The assay sample and buffer are incubated together with 1,25 (OH)2D3-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the 1,25 (OH)2D3 concentration since 1,25 (OH)2D3 from samples and 1,25 (OH)2D3-HRP conjugate compete for the anti-1,25 (OH)2D3 antibody binding site. Since the number of sites is limited, as more sites are occupied by 1,25 (OH)2D3 from the sample, fewer sites are left to bind 1,25 (OH)2D3-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The 1,25 (OH)2D3 concentration in each sample is interpolated from this standard curve. |
Quality Control on Human 1,25 dihydroxyvitamin D3 ELISA kit
| Coefficient of Variance | Intra Variation% ＜10% | |
| Inter Variation% ＜12% | ||
| Recovery | 88-118% | |
| Linearity | Diluent Ratio | Range % |
| 1:2 | 97-103 | |
| 1:4 | 92-104 | |
| 1:8 | 89-99 | |
| Specificity/Cross-reactivity | No significant cross-reactivity or interference between 1,25 (OH)2D3 and analogues was observed. | |
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Summary of the Assay Procedure for Human 1,25 dihydroxyvitamin D3 ELISA kit
