Kit for detecting G119S mutation of culex acetylcholinesterase
A technology of acetylcholinesterase and kits, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as troubles in daily life and detection troubles in daily life, and achieve the effect of simple operation
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Embodiment 1
[0042] Embodiment 1, the composition of kit
[0043] The kit consists of protein extraction solution, chromogenic reagent, inhibitor I, inhibitor II and substrate tubes.
[0044] Protein extract (1 bottle): composed of 250mM pH7.0PB buffer and Triton X100, the concentration of Triton X100 is 1% (percentage by weight); said 250mM pH7.0PB buffer consists of water, sodium dihydrogen phosphate and hydrogen phosphate disodium composition.
[0045] Chromogenic reagent (1 bottle): composed of 25mM pH7.0PB buffer, DTNB and NaHCO 3 Composition, the concentration of DTNB is 0.2mM, NaHCO 3 The concentration is 0.35mM; the 25mM pH7.0PB buffer is composed of water, sodium dihydrogen phosphate and disodium hydrogen phosphate.
[0046] Inhibitor I (1 bottle): composed of propoxur and ethanol, the concentration of propoxur is 10 -3 M.
[0047] Inhibitor II (1 bottle): composed of propoxur and ethanol, the concentration of propoxur is 10 -1 M.
[0048] Substrate tubes (20 pieces): 1.5ml...
Embodiment 2
[0051] Embodiment 2, the application of kit
[0052] 1. Using the kit prepared in Example 1 to detect the SR strain of Culex mosquito
[0053] 1. Put a single mosquito (Culex spp. SR strain, adult) in a 1.5ml centrifuge tube, add 400ul protein extract, grind the mosquito thoroughly, centrifuge at 10,000rpm for 10min at 4°C, and take the supernatant as the protein to be tested Samples (keep carefully on ice for later use).
[0054] 2. Take a substrate tube, centrifuge a little to make the drug gather at the bottom of the tube, take 1ml of chromogenic reagent and add it to the substrate tube, shake well to get the substrate (put it on ice for later use).
[0055] 3. Prepare three 0.5ml centrifuge tubes (tube No. 1, tube 2 and tube 3), add 100 μl protein sample to be tested in each tube; then add 10 μl absolute ethanol to tube 1, and add 10 μl inhibitor I to tube 2 , Add 10 μl Inhibitor II to No. 3 tube, and place it at room temperature for 15 minutes.
[0056]4. Add 80 μl of ...
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