Method for preparing protein crystal
A protein and crystal technology, applied in the field of protein crystallization, can solve the problems of complicated operation steps and high reagent requirements, and achieve the effects of high single crystal diffraction intensity, good reproducibility of results, and relaxation of crystallization conditions.
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Embodiment 1
[0023] Embodiment 1, the preparation of lysozyme crystal
[0024] (1) First prepare a NaCl storage solution with a mass percentage content of 20%, a 0.6M NaOAc buffer solution (pH is 4.5) storage solution and a 30 mg / ml lysozyme aqueous solution storage solution.
[0025] (2) Prepare 600 μL of bottom solution from the NaCl storage solution and NaOAc buffer solution prepared in step (1), its pH is 5.0, wherein the mass percentage of NaCl is 6.5%, and the molar concentration of NaOAc is 0.1M,
[0026] (3) Preparation of crystallization liquid: 2 μ L bottom solution prepared by step (2), 2 μ L 30 mg / ml lysozyme aqueous solution and 1 μ L mass percent content are respectively 1.8%, 2.4%, 3.2%, 3.6% and 7.2%, The polyvinylpyrrolidone aqueous solution with a number average molecular weight of 58,000 is composed; using the hanging drop method, the above crystallization solution is placed on a plastic cover glass, and then the crystallization drop is inverted on a 24-hole crystallizat...
Embodiment 2
[0029] Embodiment 2, the preparation of lysozyme crystal
[0030] The preparation process was the same as in Example 1, except that 1 μL of polyvinylpyrrolidone aqueous solution in the crystallization solution in Example 1 was changed to 1 μL of 1.0% by mass polyacrylic acid (number average molecular weight: 100,000) aqueous solution. Gained lysozyme crystals have a concave morphology of four adjacent (101) faces, such as figure 2 shown.
Embodiment 3
[0031] Embodiment 3, the preparation of lysozyme crystal
[0032] The preparation process is the same as in Example 1, except that 1 μL of polyvinylpyrrolidone aqueous solution in the crystallization solution in Example 1 is changed into 1 μL of ethylene oxide / oxygen An aqueous solution of a propylene diblock polymer, wherein the number average molecular weight of the ethylene oxide / propylene oxide diblock polymer is 100,000 (the mass ratio of the two blocks is 1:1). The obtained lysozyme crystals are long prisms or rods, such as image 3 shown.
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