Homocysteine kit based on aptamer fluorescence probe HCy2 and detection method thereof
A homocysteine and fluorescent probe technology, applied in the field of homocysteine HCy kits, can solve the problems of operation technical error, poor repeatability, and high test cost, and achieve high repeatability, convenient transportation, Sensitive effect
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Embodiment 1
[0034] After dissolving and preparing the required reagent components according to Example 1 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:
[0035] (1) Lysis of blood samples: Mix heparin anticoagulated whole blood and erythrocyte lysate at a volume ratio of 1:0.5, let stand for 30 minutes, then centrifuge at a medium speed for 7 minutes, and collect the supernatant;
[0036] (2) Mixed egg incubation: take 20 μl of the supernatant obtained in step 1) and 45ul of the homocysteine nucleic acid aptamer fluorescent probe reagent obtained by dissolving the homocysteine nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer Mix and incubate the eggs at room temperature for 5 minutes to obtain the test solution;
[0037] (3) Fluorescence detection: 50ul of the test solution obtained in the detecti...
Embodiment 2
[0041] After dissolving and preparing the required reagent components according to Example 2 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:
[0042] (1) Lysis of blood samples: Mix peripheral blood and erythrocyte lysate at a volume ratio of 1:2.5, let stand for 5 minutes, then centrifuge at a medium speed for 6 minutes, and collect the supernatant;
[0043] (2) Mixed egg incubation: take 50 μl of the supernatant obtained in step 1) and 30ul of the homocysteine nucleic acid aptamer fluorescent probe reagent obtained by dissolving the homocysteine nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer Mix and incubate the eggs at room temperature for 6 minutes to obtain the test solution;
[0044] (3) Fluorescence detection: 50ul of the test solution obtained in the detection step 2) of the f...
Embodiment 3
[0047] After dissolving and preparing the required reagent components according to Example 3 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:
[0048] (1) Lysis of blood samples: Mix peripheral blood and erythrocyte lysate at a volume ratio of 1:3.5, let stand for 10 minutes, then centrifuge at a medium speed for 5 minutes, and collect the supernatant;
[0049] (2) Mixed egg incubation: take 75 μl of the supernatant obtained in step 1) and 45ul of the homocysteine nucleic acid aptamer fluorescent probe reagent obtained by dissolving the homocysteine nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer Mix and incubate the eggs at room temperature for 7 minutes to obtain the test solution;
[0050] (3) Fluorescence detection: 50ul of the test solution obtained in the detection step 2) of the ...
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