Protocol tips
| Upstream tips |
Protocol tips |
Downstream tips |
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For expression in E. coli, the constructs were transformed into TP1000 (ΔmobAB) cells (Palmer et al., 1996). E. coli were grown at 30°C in Luria-Bertani medium supplemented with 150 μg/ml ampicillin, 1 mM molybdate, and 20 μM isopropyl β-d-thiogalactoside. |
Cells were harvested by centrifugation after 24 h of growth and resuspended in 50 mM sodium phosphate buffer (pH 8.0) containing 300 mM NaCl and frozen at −20°C until purification. |
| Protocol tips |
| For expression in E. coli, the constructs were transformed into TP1000 (ΔmobAB) cells (Palmer et al., 1996). E. coli were grown at 30°C in Luria-Bertani medium supplemented with 150 μg/ml ampicillin, 1 mM molybdate, and 20 μM isopropyl β-d-thiogalactoside. |
| Downstream tips |
| Cells were harvested by centrifugation after 24 h of growth and resuspended in 50 mM sodium phosphate buffer (pH 8.0) containing 300 mM NaCl and frozen at −20°C until purification. |
Publication protocol
For expression in E. coli, the constructs were transformed into TP1000 (ΔmobAB) cells (Palmer et al., 1996). E. coli were grown at 30°C in Luria-Bertani medium supplemented with 150 μg/ml ampicillin, 1 mM molybdate, and 20 μM isopropyl β-d-thiogalactoside. Cells were harvested by centrifugation after 24 h of growth and resuspended in 50 mM sodium phosphate buffer (pH 8.0) containing 300 mM NaCl and frozen at −20°C until purification.
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