Application of HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) in xanthine oxidase inhibitory peptide
A technology of hydroxyethylpiperazine ethanesulfonic acid and xanthine oxidase, which can be used in peptide/protein components, medical preparations containing active ingredients, organic active ingredients, etc. and other problems, to achieve the effect of enhancing the activity and improving the inhibition rate of xanthine oxidase
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Embodiment 1
[0028] The enhancement effect of 4-hydroxyethylpiperazineethanesulfonic acid on tuna peptide xanthine oxidase inhibitors, the specific experimental steps are as follows:
[0029] (1) Prepare 150mM HEPES, Tris-HCl, PB buffer solution with pH value of 7.4 at 37°C, store at 4°C until use;
[0030] (2) Prepare the required sample (tuna peptide) with the above-mentioned 150mM buffer solution, and the sample concentration is 40mg / ml;
[0031] (3) Add 50 μL tuna peptide or 50 μL buffer (blank) and 150 μL xanthine to the 96-well ELISA plate in turn, make 3 parallels for each sample, add 50 μL xanthine oxidase after incubation at 37°C for 5 minutes, and read once every 30 seconds Absorbance value, a total of 50 times 25min. After reading, stop the reaction with 80 μL of HCl, take the reaction solution and dilute it 10 times with the corresponding buffer solution, pass it through a 0.25 μm water-based membrane, and measure the concentration of uric acid;
[0032] (4) high performance ...
Embodiment 2
[0039] The enhancement effect of 4-hydroxyethylpiperazineethanesulfonic acid on saury peptide xanthine oxidase inhibitor, the specific experimental steps are as follows:
[0040] (1) Prepare 150mM HEPES, Tris-HCl, PB buffer solution with pH value of 7.4 at 37°C, store at 4°C until use;
[0041] (2) Prepare the required sample (saury peptide) with the above-mentioned 150mM buffer solution, and the sample concentration is 40mg / ml;
[0042] (3) Add 50 μL saury peptide or 50 μL buffer solution (blank) and 150 μL xanthine to the 96-well ELISA plate in sequence, make 3 parallels for each sample, add 50 μL xanthine oxidase after incubation at 37°C for 5 minutes, and read for 30 seconds One absorbance value, a total of 50 times 25min. After reading, stop the reaction with 80 μL of HCl, take the reaction solution and dilute it 10 times with the corresponding buffer solution, pass it through a 0.25 μm water-based membrane, and measure the concentration of uric acid;
[0043] (4) high ...
Embodiment 3
[0050] The enhancement effect of 4-hydroxyethylpiperazineethanesulfonic acid on xanthine oxidase inhibitor of walnut meal protein peptide, the specific experimental steps are as follows:
[0051] (1) Prepare 150mM HEPES, Tris-HCl, PB buffer solution with pH value of 7.4 at 37°C, store at 4°C until use;
[0052] (2) Prepare the required sample (walnut meal protein peptide) with the above-mentioned 150mM buffer solution, and the sample concentration is 40mg / ml;
[0053] (3) Add 50 μL walnut meal protein peptide or 50 μL buffer solution (blank) and 150 μL xanthine to the 96-well ELISA plate in sequence, make 3 parallels for each sample, keep warm at 37°C for 5 minutes, then add 50 μL xanthine oxidase for 30 seconds Read the absorbance value once, a total of 50 times for 25 minutes. After reading, stop the reaction with 80 μ LHCl, take the reaction solution and dilute it 10 times with the corresponding buffer solution, pass it through a 0.25 μm water-based membrane, and measure t...
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