Protein extraction method and application
A protein and the resulting technology, applied in the biological field, can solve the problems of less total protein extraction, toxic to human health, and difficulty in meeting demand, and achieve stable dewaxing effect, not easy to be damaged and broken, simple and time-saving dewaxing steps Effect
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Embodiment 1
[0074] A method for extracting protein from paraffin-embedded tissue, specifically comprising the steps of:
[0075] (1) Remove excess paraffin from the paraffin-embedded tissue block, cut it into tissue sections with a thickness of 28 μm, and then place it in a nested screen chamber (such as figure 2 shown);
[0076] (2) immerse the section described in step (1) into the TO transparent agent for dewaxing, and perform dewaxing treatment 3 times, soaking for 10 minutes each time;
[0077] (3) Ethanol gradient hydration treatment was performed on the tissue slices obtained in step (2). Firstly, the tissue slices were hydrated twice by immersing in absolute ethanol for 5 minutes each time, then immersed in 95% ethanol for 5 minutes, and then immersed in 75% ethanol for 5 minutes. Finally, immerse in distilled water twice, each time for 5 minutes;
[0078] (4) Drain the tissue slices obtained in step (3), collect them in a weighed 1.5mL EP tube, wash the tissue slices twice wit...
Embodiment 2
[0083] (1) Remove excess paraffin from the paraffin-embedded tissue block, cut it into tissue sections with a thickness of 20 μm, and then place it in a nested screen chamber;
[0084] (2) immerse the tissue section described in step (1) into TO transparent agent for dewaxing, and perform dewaxing treatment 4 times, soaking for 12 minutes each time;
[0085] (3) Carry out ethanol gradient hydration treatment to the tissue slices obtained in step (2), first immerse the tissue slices in absolute ethanol for hydration 3 times, each time for 5 min, then immerse in 95% ethanol for 5 min, then immerse in 75% ethanol for 5 min, Finally, immerse in distilled water 3 times, each time for 4 minutes;
[0086] (4) Drain the tissue slices obtained in step (3), collect them in a weighed 1.5mL EP tube, wash the tissue slices twice with PBS, each time for 8 min, centrifuge at 14000 rpm for 5 min, discard the supernatant, and Calculation of the weight of the slice by the method of measurement...
Embodiment 3
[0091](1) Remove excess paraffin from the paraffin-embedded tissue block, cut it into tissue sections with a thickness of 10 μm, and then place it in a nested screen chamber;
[0092] (2) immerse the tissue section described in step (1) into TO transparent agent for dewaxing, dewaxing treatment 2 times, soaking for 15min each time;
[0093] (3) Ethanol gradient hydration treatment was performed on the tissue slices obtained in step (2). Firstly, the tissue slices were hydrated once by immersing in absolute ethanol for 8 min each time, then immersed in 95% ethanol for 3 min, and then immersed in 75% ethanol for 5 min. Finally, immerse in distilled water 3 times, each time for 3 minutes;
[0094] (4) Drain the tissue slices obtained in step (3), collect them in a weighed 1.5mL EP tube, wash the tissue slices once with PBS, 10 min each time, centrifuge at 13,000 rpm for 5 min, discard the supernatant, and Calculation of the weight of the slice by the method of measurement;
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