Single sperm freezing carrier in combination with monoclonal antibodies and freezing and unfreezing method
A monoclonal antibody and freezing carrier technology, which can be used in measurement devices, instruments, scientific instruments, etc., and can solve problems such as difficulty in finding sperm.
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Embodiment 1
[0031] The monoclonal antibody in this example binds to the single sperm cryocarrier, the monoclonal antibody is ACRV1acrosomal vesicle protein 1 [Homo sapiens (human)], the manufacturer of the monoclonal antibody is sant cruz, the article number is sc5163.
[0032] The volume ratio of monoclonal antibody to sperm is (10-1000):1 to form antibody sperm solution.
[0033] The freezing and thawing method of the monoclonal antibody combined with the single sperm freezing carrier carried out according to the following steps:
[0034] (1) Add the slides one by one into the piranha solution and soak for 15 minutes, then put them into pure water for ultrasonication, change the water at least three times to remove the organic matter on the surface of the slides, and dry the slides in a drying oven at 105°C for 30 minutes. spare;
[0035] (2) Dissolving 3-aminopropyltriethoxysilane (APTES) in absolute ethanol with a mass ratio of 2.5%, soaking the above-mentioned oven-dried slides in t...
Embodiment 2
[0043] The monoclonal antibody of this example is combined with a single sperm freezing carrier, the monoclonal antibody is PH-20 [Homosapiens], the manufacturer of the monoclonal antibody is sant cruz, and the article number is sc5163. The volume ratio of monoclonal antibody to sperm is (10-1000):1 to form antibody sperm solution.
[0044] The freezing and thawing method of the monoclonal antibody combined with the single sperm freezing carrier carried out according to the following steps:
[0045] (1) Add the slides one by one into the piranha solution and soak for 150 minutes, then put them into pure water for ultrasonication, change the water at least three times to remove the organic matter on the surface of the slides, and dry the slides in a drying oven at 80°C for 30 minutes. spare;
[0046] (2) Dissolving 3-aminopropyltriethoxysilane (APTES) in absolute ethanol with a mass ratio of 10%, soaking the above-mentioned oven-dried slides in the liquid for 72 hours;
[004...
Embodiment 3
[0055] The monoclonal antibody in this example binds to the single sperm cryocarrier, the monoclonal antibody is ACRV1acrosomal vesicle protein 1 [Homo sapiens (human)], the manufacturer of the monoclonal antibody is sant cruz, the article number is sc5163. The volume ratio of monoclonal antibody to sperm is (10-1000):1 to form antibody sperm solution.
[0056] The freezing and thawing method of the monoclonal antibody combined with the single sperm freezing carrier carried out according to the following steps:
[0057] (1) Add the slides one by one into the piranha solution and soak for 240 minutes, then put them into pure water for ultrasonication, change the water at least three times to remove the organic matter on the surface of the slides, and dry the slides in a drying oven at 150°C for 30 minutes. spare;
[0058] (2) Dissolving 3-aminopropyltriethoxysilane (APTES) in absolute ethanol with a mass ratio of 1%, soaking the above-mentioned oven-dried glass slides in the l...
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