I have cloned pET28a vector with BF AchE gene, the recombinant pET 28a vector is transformed in BL21 cells and for purifying His-tagged BF AchE enzyme, Please suggest the lysis buffer that I can use, so that the enzymatic activity of the enzyme is not hampered.
Manik Goel
You can try the sonication method for lysis after resuspending in a lysis buffer containing lysozyme, and the lysis buffer should depend on your needs. It can include components like Tris, HEPES, having buffer capacity, PMSF for stability, Lysozyme, EDTA if it's not a metal-bound protein, NaCl and non-ionic detergent like Triton X-100.
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