Method for separating heat shock protein 60 from nematocyst venom of Cyanea nozakii

Method for separating heat shock protein 60 from nematocyst venom of Cyanea nozakii

  • CN 102,153,639 A
  • Filed: 12/10/2010
  • Published: 08/17/2011
  • Est. Priority Date: 12/10/2010
  • Status: Active Application
First Claim
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1. method of separating heat shock protein 60 from rosy clouds jellyfish stinging capsule toxin is characterized in that:

  • The rosy clouds jellyfish stinging capsule cell that

         1) will obtain from rosy clouds jellyfish tentacle adds in the precooling damping fluid broken, and broken back low-temperature centrifugation is collected supernatant liquor, and is stand-by;

    2) step

         1) gained supernatant liquor be rosy clouds jellyfish stinging capsule cytotoxin under 2-6 ℃

    to pH7.820mM Tris-HCl damping fluid dialysed overnight, low-temperature centrifugation is then collected supernatant liquor, and is stand-by;

    3) with step

         2) the gained supernatant liquid filtering, then the anionresin tree post that contains DEAE SepharoseFast Flow on it is separated, at first adopt the pH7.820mMTris-HCl buffer solution elution, then adopt and contain the pH7.820mM Tris-HCl damping fluid that concentration is respectively the NaCl of 0.1-2M different concns and carry out gradient elution, collect each elution peak, be that the ultrafiltration pipe of 3kDa concentrates then with molecular weight cut-off, stand-by;

    4) be the spissated enriched material of the ultrafiltration pipe gel resin Superdex75 column purification separation of 3kDa with the step

         3) molecular weight cut-off, adopt and contain the pH7.820mM Tris-HCl buffer solution elution that concentration is the NaCl of 0.15-0.5M that flow velocity is 0.2-0.5mLmin -1, collect each elution peak, wherein the component in the efflux volume 50mL-80mL is heat shock protein 60.

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