Use of trypsin
A technology for trypsin and uses, applied in the field of trypsin, can solve problems such as toxic effects, destruction of cell components, etc.
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Embodiment 1
[0025] In this example, the application of trypsin in scavenging free radicals.
[0026] 1. Materials and methods
[0027] 1. Materials
[0028] Escherichia coli wild-type strain MG1655 was donated by Professor Imlay of Illinois State University. Streak inoculate the dry powder of the above-mentioned strains stored in freeze-dry on LB plates, culture at 37°C for more than 16 hours, pick a single colony into the LB liquid medium, and cultivate at 37°C until the concentration of the bacterial solution is about 10 9 cells / ml, glycerol -20 ℃ to save the strains, for later use.
[0029] 2. Trypsin treatment
[0030] Trypsin (500U / mg) was purchased from Amersco, take 100mg.ml -1 Trypsin was added to O 2 ·- Incubate the resulting mixed system at 37°C for 30min, then add 25μl of 10mg.ml -1 Soybean trypsin inhibitor terminates the reaction.
[0031] 3. Sample processing
[0032] Sample with 0.5mM Cu 2+ Either incubate with 25mM EDTA for 0.5h or incubate with 1mM DDC at 28°C for...
Embodiment 2
[0053] The application of trypsin in anti-aging (delaying aging) in this example.
[0054] The tumor mice with no significant difference in the number of voluntary activities were selected and divided into two groups, 10 in each group. The control group was intragastrically given 10 mL / kg of normal saline according to the weight of the mice, and the experimental group was given intravenous injection of normal saline (10 mL / kg) containing 0.6 mg / mL trypsin according to the weight of the mice, and the survival time of the mice was recorded. The results showed that the mice in the control group died within 1 week, and the survival period of the mice in the trypsin treatment group was extended to 8 weeks.
Embodiment 3
[0056] In this example, the application of trypsin in anti-tumor (inhibition of tumor cell proliferation activity).
[0057] Hela cells in the logarithmic growth phase were collected, and the experimental group was treated with trypsin (0.6 mg / ml), and the control group was added with 200 μL of bovine serum culture medium, and the temperature was 5% CO at 37°C. 2 and fully saturated humidity CO 2 Cultivate in the incubator for 24-48 hours, and detect the cell survival rate by MTT method. The results showed a 30 percent decrease in tumor cell survival in the trypsin-treated group.
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