Author: Bouvette, Jonathan; Korkut, Dursun Nizam; Fouillen, Aurélien; Amellah, Soumiya; Nanci, Antonio; Durocher, Yves; Omichinski, James G.; Legault, Pascale
Title: High-yield production of human Dicer by transfection of human HEK293-EBNA1 cells grown in suspension Document date: 2018_12_6
ID: 012ipcdr_7
Snippet: An efficient multi-step purification protocol was developed to maximize purification yields (Fig. 2b) . While optimizing this protocol, we noticed that Dicer precipitated either at high concentration or when in contact with metallic surfaces such as stainless steel needles, FPLC titanium alloy pump heads and the non-reactive Hastelloy sample cell of the isothermal titration calorimeter. Thus, the final purification protocol was optimized to avoid.....
Document: An efficient multi-step purification protocol was developed to maximize purification yields (Fig. 2b) . While optimizing this protocol, we noticed that Dicer precipitated either at high concentration or when in contact with metallic surfaces such as stainless steel needles, FPLC titanium alloy pump heads and the non-reactive Hastelloy sample cell of the isothermal titration calorimeter. Thus, the final purification protocol was optimized to avoid protein contact with metallic surfaces and minimize sample handling that could lead to protein aggregation and precipitation.
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