Diacetyl spermine detection kit and preparation method and application thereof
A technology of diacetylspermine and detection kits, which is applied in the field of biological detection, can solve the problems of elevated tumor incidence, missed treatment time, etc., and achieve the effect of simple operation
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Embodiment 1
[0044] Kit preparation:
[0045] ①Take 1ml of colloidal gold, add 7μL of 0.2M K 2 CO 3 After mixing, add 20 μg DiAcSpm monoclonal antibody, mix and let stand for 10 minutes, add 10 μL 20% BSA to block, mix again and let stand for 10 minutes, centrifuge for 7 minutes (9000r), discard the supernatant, and redissolve the precipitate with 1000 μL reconstitution solution;
[0046] ②Take 1ml of colloidal gold, add 2-10μL of 0.2M K 2 CO 3 After mixing, add 10-30 μg CRE monoclonal antibody, mix and let stand for 10 minutes, add 10 μL 20% BSA to block, mix again and let stand for 10 minutes, centrifuge for 7 minutes (9000r), discard the supernatant, and redissolve the precipitate with 1000 μL reconstitution solution;
[0047] ③Take 2ml of colloidal gold, add 10μL of 0.2M K 2 CO 3 After mixing, add 30 μg rabbit antibody, mix and let stand for 10 minutes, add 20 μL 20% BSA to block, mix again and let stand for 10 minutes, centrifuge for 7 minutes (9000r), discard the supernatant, an...
Embodiment 2
[0053] Kit preparation:
[0054] ①Take 1ml of colloidal gold, add 7μL of 0.2M K 2 CO 3 After mixing, add 20 μg DiAcSpm monoclonal antibody, mix and let stand for 10 minutes, add 10 μL 20% BSA to block, mix again and let stand for 10 minutes, centrifuge for 7 minutes (9000r), discard the supernatant, and redissolve the precipitate with 1000 μL reconstitution solution;
[0055] ②Take 1ml of colloidal gold, add 2-10μL of 0.2M K 2 CO 3 After mixing, add 10-30 μg CRE monoclonal antibody, mix and let stand for 10 minutes, add 10 μL 20% BSA to block, mix again and let stand for 10 minutes, centrifuge for 7 minutes (9000r), discard the supernatant, and redissolve the precipitate with 1000 μL reconstitution solution;
[0056] ③Take 2ml of colloidal gold, add 10μL of 0.2M K 2 CO 3 After mixing, add 30 μg rabbit antibody, mix and let stand for 10 minutes, add 20 μL 20% BSA to block, mix again and let stand for 10 minutes, centrifuge for 7 minutes (9000r), discard the supernatant, an...
Embodiment 3
[0062] The kit obtained above was tested, and a total of 25 healthy samples, 25 samples of rectal cancer patients, and 26 samples of gastric cancer patients were detected, and the sample volume was 80 μL. Product curve (see Figure 8) and product application test statistics (see Figure 9 and Table 1).
[0063] Table 1
[0064]
[0065] Carry out product curve by the detection result of above-mentioned Fig. 8, pass Y=A+C / [1+exp{-B(X-D)}] and Y=(A-D) / [1+(X / C)^B]+ D fitting gives R 2 ≥0.9801 by R 2 It can be seen that each index detected by the obtained kit of the present invention meets the requirements for use of the product.
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