Zebra fish vitellogenin immunoblotting reagent kit as well as detection method and application thereof
A technology of western blotting and original protein, which is applied in the direction of measuring devices, biological tests, material inspection products, etc., can solve the problems of affecting effect, poor specificity and sensitivity, etc., achieve large induction amount, simplify experimental methods, and reduce induction time and workload effects
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[0021] The preparation of the western blot kit for detection of zebrafish vitellogenin is divided into the following steps:
[0022] (1) Induction and purification of zebrafish vitellogenin
[0023] Preparation of pure zebrafish vitellogenin: zebrafish were induced to produce vitellogenin by exposing water to 17β-estradiol, and the zebrafish were placed in a 2L glass jar, and 10 zebrafish were placed in each jar, 17β-estradiol The exposure concentration of estradiol was 200 μg / L, all the water was changed every day, and the corresponding 17β-estradiol was added again, and the feed was fed in the morning and evening, and the water temperature was 26°C; After the zebrafish is anesthetized, take it out and weigh it, add 3 times the volume of 4°C pre-cooled PBS buffer (containing 1mM PMSF, pH 7.5), homogenize with a glass homogenizer under ice bath conditions, and centrifuge at 4°C at 8000g After 10 minutes, the supernatant was collected; all the collected supernatants were used ...
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