E03O0874 Mouse 25 Hydroxyvitamin D3 ELISA kit
The Mouse 25 Hydroxyvitamin D3 ELISA kit can be used to identify samples from the mouse species. 25 Hydroxyvitamin D3 can also be called 25-Hydroxyvitamin D3, 25-Hydroxycholecalciferol, Calcidiol, 25(OH)D3.
Product Information | |
Cat. No. | E03O0874 |
Product Name | Mouse 25 Hydroxyvitamin D3 ELISA kit |
Species | Mouse |
Product Size | 48 Tests / 96 Tests |
Concentration | 5.0-100ng/ml |
Sensitivity | 1.0 ng/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 ng/mL | 1 vial |
STANDARD B (0.5mL) | 5.0 ng/mL | 1 vial |
STANDARD C (0.5mL) | 10 ng/mL | 1 vial |
STANDARD D (0.5mL) | 25 ng/mL | 1 vial |
STANDARD E (0.5mL) | 50 ng/mL | 1 vial |
STANDARD F (0.5mL) | 100 ng/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 |
25(OH)D3 ELISA kit applies the competitive enzyme immunoassay technique utilizing an anti-25(OH)D3 antibody and an 25(OH)D3-HRP conjugate. The assay sample and buffer are incubated together with 25(OH)D3-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 25(OH)D3 concentration since 25(OH)D3 from samples and 25(OH)D3-HRP conjugate compete for the anti-25(OH)D3 antibody binding site. Since the number of sites is limited, as more sites are occupied by 25(OH)D3 from the sample, fewer sites are left to bind 25(OH)D3-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The 25(OH)D3 concentration in each sample is interpolated from this standard curve. |
Coefficient of Variance | Intra Variation% ＜10% | |
Inter Variation% ＜12% | ||
Recovery | 95-102% | |
Linearity | Diluent Ratio | Range % |
1:2 | 93-105 | |
1:4 | 88-106 | |
1:8 | 86-108 | |
Specificity/Cross-reactivity | No significant cross-reactivity or interference between 25(OH)D3 and analogues was observed. | |
E03O0874 has been referenced in the below publications：
In vitro vitamin K2 and 1α, 25-dihydroxyvitamin D3 combination enhances osteoblasts anabolism of diabetic mice.
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