Dissociation method of 25-hydroxyvitamin D binding protein
A hydroxyvitamin and binding protein technology, applied in the field of vitamin D detection, can solve the problems of poor dissociation effect, toxicity, poor stability, etc., and achieve the effects of improving detection results, strong reducibility and improving stability
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Embodiment 1
[0020] The operator prepares mother liquor in advance, selects dithiothreitol with a concentration of 0.01g / ml, mixes dithiothreitol with water to form a 1mol / L dithiothreitol aqueous solution, and configures multiple mother liquors, each with a capacity of 1ml, the prepared mother solution is stored for later use, the spare mother solution is stored at -20°C for short-term use, and the mother solution is stored at -80°C for long-term storage; take 200 μL serum sample, add 20 μL Mother liquor, vortex 1min to mix, then put the mixture into a centrifuge for 5min, centrifuge at 13000rpm, temperature 4°C, centrifuge for 10min under this condition; take the supernatant of the obtained solution, add 200μL ethanol , manually vortexed for 1min to mix, and the mixed solution was left to stand for 5min; the obtained solution was put into a centrifuge with a centrifugal speed of 13000rpm and a temperature of 4°C, and centrifuged for 10min under these conditions. After the centrifugation w...
Embodiment 2
[0022] The operator prepares mother liquor in advance, selects dithiothreitol with a concentration of 0.015g / ml, mixes dithiothreitol with water to form a 1mol / L dithiothreitol aqueous solution, and configures multiple mother liquors, each with a capacity of 1ml, the prepared mother solution is stored for later use, the spare mother solution is stored at -20°C for short-term use, and the mother solution is stored at -80°C for long-term storage; take 250 μL of serum samples, add 25 μL of mother solution , vortexed for 1 min to mix, and then put the mixture into a centrifuge for 5 min to centrifuge at a speed of 13000 rpm and a temperature of 4 °C for 10 min under these conditions; take the supernatant of the obtained solution, add 250 μL of ethanol, Manually vortex for 1 min to mix well, then let the mixed solution stand for 5 min; put the obtained solution into a centrifuge with a centrifugal speed of 13,000 rpm and a temperature of 4°C, and centrifuge for 10 min under these co...
Embodiment 3
[0024] The operator prepares the mother liquor in advance, selects dithiothreitol with a concentration of 0.03g / ml, mixes dithiothreitol with water to form a 1mol / L aqueous solution of dithiothreitol, configures multiple mother liquors, and the capacity of each mother liquor is 1ml, the prepared mother solution is stored for later use, the spare mother solution is stored at -20°C for short-term use, and the mother solution is stored at -80°C for long-term storage; take 300 μL serum sample, add 30 μL Mother liquor, vortex 1min to mix, then let the mixture stand for 5min, put it into a centrifuge, centrifuge at 13000rpm, temperature 4°C, centrifuge for 10min under this condition; take the supernatant of the obtained solution, add 300μL ethanol , manually vortexed for 1min to mix, and the mixed solution was left to stand for 5min; the obtained solution was put into a centrifuge with a centrifugal speed of 13000rpm and a temperature of 4°C, and centrifuged for 10min under these con...
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