Natural antioxidant peptide Macrotympanain-Al and preparation method and application thereof
An anti-oxidative peptide and anti-oxidative activity technology, which can be used in the preparation methods of peptides, chemical instruments and methods, cosmetic preparations, etc., and can solve the problems of limited oxidative activity and high price.
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[0020] ① Acquisition of skin secretions: Stimulate the skin of the Yunnan stinky frog with a voltage of 6V for 6s, then wash the body of the stinky frog with 25mM Tris-HCl buffer (pH=7.8), collect the skin secretions, and put the collected secretions in Centrifuge at 4000rpm for 15min, vacuum freeze-dry the supernatant, and store at -80°C for later use;
[0021] ②Purification of polypeptide: redissolve the lyophilized supernatant sample, put it on a Sephadex G-75 gel filtration column equilibrated with 25mM Tris-HCl buffer (pH=7.8, containing 0.1M NaCl) in advance, and use 0.3ml / min flow rate for elution, collected every 10min. The eluted product was monitored at 280nm, the separation and purification map was drawn, and the antioxidant active ingredients were collected, and used C 4 The column was freeze-dried after removing the salt inside. Afterwards, the sample was dissolved with 20 mM NaAc-HAc buffer (pH=4.0), purified with a Resource S separation column equilibrated wi...
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