purification:tev:c_protocol
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| purification:tev:c_protocol [2016/07/18 08:31] – created bonnefon | purification:tev:c_protocol [2023/11/01 20:19] (current) – external edit 127.0.0.1 | ||
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| * isopropyl β-D-thiogalactopyranoside (IPTG) (0.5 mM) induction. | * isopropyl β-D-thiogalactopyranoside (IPTG) (0.5 mM) induction. | ||
| * The cells were then lysed using sonication. | * 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, | + | * 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, |
| - | * The protein was eluted with Buffer B((25 mM sodium phosphate (pH 8.0), 10% (v/v) glycerol, | + | * The protein was eluted with Buffer B((25 mM sodium phosphate (pH 8.0), 10% (v/v) glycerol, |
| - | * The eluted protein was then applied to a Superdex 75 (16/10) column in Buffer C((25 mM Na2PO4 | + | * The eluted protein was then applied to a Superdex 75 (16/10) column in Buffer C((25 mM sodium phosphate |
| * The eluted fractions containing TEV were pooled, flash-frozen in liquid nitrogen, and stored at −80°C. | * The eluted fractions containing TEV were pooled, flash-frozen in liquid nitrogen, and stored at −80°C. | ||
purification/tev/c_protocol.1468830696.txt.gz · Last modified: (external edit)
