lysozymeによる溶菌や、界面活性剤による溶菌も比較検討すべきです。
例えば、Sigma社のLyticBは自作できます。
CelLytic B & CelLytic 10× Lysis Reagent”
This example sets forth results that compare a zwitterionic detergent combination of the present invention with a commercially available lysis solution, CelLytic B (Sigma-Aldrich Co. Product No. B3553). This reagent contains 1% (w/v) octyl-β-D-thioglucopyranoside, with a Tris buffer, 40 mM Trizma® HCl(pH 8.0). The soluble protein fraction is extracted using a detergent lysis solution, and then affinity purified using HIS-Select Nickel Spin Columns (Sigma-Aldrich Co. Product No. H7787).
Preparation of Lysis Solutions. The zwitterionic detergent combination of the current invention was prepared by dissolving 10 grams of 3-(N,N-Dimethyltetradecylammonio)propanesulfonate (Sigma-Aldrich Co. Product No. T7763) and 1 gram of C7BzO (Sigma-Aldrich Co. Product No. C0856) into 100 ml of ultrapure water (“CelLytic 10× Lysis Reagent”). The “Tris Working Reagent” was prepared by diluting 1 ml of the CelLytic 10× Lysis Reagent into 8.6 ml of ultrapure water and 400 μl of 1M Tris solution. The “Phosphate Working Reagent” was prepared by diluting 1 ml of the CelLytic 10× Lysis Reagent into 5 ml of 2× Saline Solution (600 mM NaCl, 100 mM sodium phosphate, pH 8.0) and 4 ml of ultrapure water. |
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