Liquid 25-hydroxyvitamin D calibrator diluent
A hydroxyvitamin and calibrator technology, applied in biological testing, material testing products, preparation of samples for testing, etc., can solve the problems of increasing production costs of enterprises, difficult preservation of calibrators, deviation of test results, etc. The effect of production cycle and stability guarantee
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Embodiment 1
[0016] Example 1 Preparation of liquid 25-hydroxyvitamin D calibrator dilution
[0017] Weigh a certain amount of 7-dehydrocholesterol, then dissolve it in a small amount of absolute ethanol, then weigh a certain proportion of human serum albumin and Proclin 300, add it to 100mmol / L PBS buffer, stir for 30min, that is Prepare a 25-hydroxyvitamin D calibrator dilution, which contains human serum albumin 3% w / v, Proclin 300 0.2% w / v, and 7-dehydrocholesterol 2.0mmol / L.
Embodiment 2
[0018] Example 2 Preparation of relevant reagents for the detection of 25-hydroxyvitamin D content in human serum samples
[0019] 1. Preparation of 25-hydroxyvitamin D series calibrator
[0020] Take a certain amount of 25-hydroxyvitamin D antigen, add respectively to the calibration product dilution prepared in Example 1, and prepare the labeled concentration as 0ng / ml, 10ng / ml, 20ng / ml, 40ng / ml, 80ng / ml, 140ng / ml of a series of calibration products, the color of the bottle cap is white, yellow, green, blue, red, purple, black;
[0021] 2. Preparation of magnetic particle suspension coated with anti-25 hydroxyvitamin D antibody
[0022] Take an appropriate amount of carboxyl magnetic particles according to the amount used, and activate them under acidic conditions with excess EDC and NHS. The activation time is 30 minutes. After the activation is completed, add a magnetic field and let it stand for 5 minutes to separate the magnetic particles from the liquid. Discard the s...
Embodiment 3
[0029] Example 3 Detection of 25-hydroxyvitamin D content in human serum samples
[0030] The relevant reagents prepared in Example 2 are used to detect the content of 25-hydroxyvitamin D in human serum samples, and the specific steps are:
[0031] 1. Take out a certain amount of reaction containers and number them. Add 50 μl of calibration / quality control / clinical serum according to the experimental requirements;
[0032] 2. Shake the magnetic particle suspension, add 20 μl to each well;
[0033] 3. Add 50 μ of enzyme marker to each well;
[0034] 4. Mix the solution in the reaction vessel evenly and incubate at 37°C for 15 minutes;
[0035] 5. Using magnetic separation and washing equipment, wash the magnetic particles in the reaction vessel with washing liquid for 5 times;
[0036] 6. The reaction vessel after washing is fully oscillated to disperse the magnetic particles;
[0037] 7. Add 50 μl each of luminescent substrate A and luminescent substrate B to each well, s...
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