Anti-oxidation medicine screening kit and use method thereof
A kit and anti-oxidation technology, applied in the field of biomedicine, can solve the problems of nematode screening models without intellectual property protection and retention, and achieve the effects of intuitive and reliable results, simple use methods, and simple and fast operations
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Embodiment 1
[0062] The assembly of embodiment 1 kit
[0063] A kind of antioxidant drug screening kit according to the present invention, the kit includes the following reagents:
[0064] ①. Reagent A: Caenorhabditis elegans suspension;
[0065] ②. Reagent B: nematode culture medium;
[0066] ③. Reagent C: Escherichia coli OP50;
[0067] ④. Reagent D: 2′,7′ dichlorofluorescein ethylene glycol;
[0068] The kit also contains a multiwell plate, which is black with Parafilm.
[0069] The porous plate is a 384-well microplate.
[0070] Wherein, the concentration of the reagent A is 50 Caenorhabditis elegans / 20 μl, the concentration of the reagent C is 10 mg Escherichia coli / ml, and the volume of the reagent B is 25 ml.
[0071] Wherein, the reagent C is a freeze-dried powder or a bacterial solution containing live Escherichia coli OP50.
[0072] The reagent B is S liquid culture medium.
[0073] Divide the above reagents and multi-well plates into suitable containers and place them in ...
Embodiment 2
[0076] Example 2 Synchronization of Caenorhabditis elegans
[0077] The Caenorhabditis elegans in the reagent A of the present invention is a nematode hatched to the L1 stage after synchronization, and the steps of Caenorhabditis elegans synchronization are as follows: wash down the Caenorhabditis elegans on the plate with M9 buffer, Inhale the washing solution into a centrifuge tube; centrifuge at 4000rpm for 3min, remove the supernatant; add the lysate; shake on a vortex instrument for 7min to rupture the worm body of Caenorhabditis elegans and release eggs; centrifuge at 4000rpm for 3min, remove the supernatant, and collect the precipitate to obtain A large number of Caenorhabditis elegans eggs; washed twice with M9 buffer, then added 1ml of M9 buffer, placed in an incubator at 20°C for 48 hours to incubate to obtain Caenorhabditis elegans hatched to the L1 stage after synchronization.
Embodiment 3
[0078] The use of embodiment 3 kit
[0079] (1) Use the Antioxidant Drug Screening Kit to screen antioxidant drugs, set a blank control group and a drug solution measurement group, and set each well in the multi-well plate as an independent experimental unit;
[0080] (2) Drug solution measurement group, add reagent B, the solution containing the drug solution to be tested, 5 μl reagent C and 20 μl reagent A to the wells of the multi-well plate in sequence under sterile conditions, so that the total volume of each well is 50 μl, set Set four parallel;
[0081] (3) Blank control group, under aseptic conditions, add reagent B, sterile water or vehicle equal to the solution of the drug solution to be tested, 5 μl reagent C and 20 μl reagent A to the wells of the multi-well plate in sequence, so that each well The total volume is 50 μl, and four parallels are set; wherein, the solution of the organic solvent is DMSO and ethanol;
[0082] (4) After adding the sample, seal the mul...
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