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--- purification:tev:c_protocol [2016/07/18 08:31] – bonnefon
+++ purification:tev:c_protocol [2023/11/01 20:19] (current) – external edit 127.0.0.1
@@ Line 9: / Line 9: @@
   * isopropyl β-D-thiogalactopyranoside (IPTG) (0.5 mM) induction.
   * The cells were then lysed using sonication.
-  * The sample was then applied to 1 mL of Ni-NTA agarose (GE Healthcare) pre-equilibrated with Buffer A((25 mM sodium phosphate (pH 8.0), 10% (v/v) glycerol, 0.2 M NaCl, and 25 mM imidazole)).
+  * The sample was then applied to 1 mL of Ni-NTA agarose (GE Healthcare) pre-equilibrated with Buffer A((25 mM sodium phosphate (pH 8.0), 10% (v/v) glycerol, 200 mM NaCl, and 25 mM imidazole)).
-  * The protein was eluted with Buffer B((25 mM sodium phosphate (pH 8.0), 10% (v/v) glycerol, 0.2 M NaCl, and 500 mM imidazole)).
+  * The protein was eluted with Buffer B((25 mM sodium phosphate (pH 8.0), 10% (v/v) glycerol, 200 mM NaCl, and 500 mM imidazole)).
-  * The eluted protein was then applied to a Superdex 75 (16/10) column in Buffer C((25 mM Na2PO4 (pH 8.0), 200 mM NaCl, 10% (v/v) glycerol, 5 mM β-mercaptoethanol)).
+  * The eluted protein was then applied to a Superdex 75 (16/10) column in Buffer C((25 mM sodium phosphate (pH 8.0), 10% (v/v) glycerol, 200 mM NaCl, and 5 mM β-mercaptoethanol)).
   * The eluted fractions containing TEV were pooled, flash-frozen in liquid nitrogen, and stored at −80°C.