User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/06: Difference between revisions
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** Cells are sonicated under 8mV for 30 seconds with ______Sonicator. | ** Cells are sonicated under 8mV for 30 seconds with ______Sonicator. | ||
** Cells are immediately placed on ice for 30 seconds | ** Cells are immediately placed on ice for 30 seconds | ||
* After | * After sonication, the E.coli cells were transferred from a 25mL falcon tube into a 30mL centrifuge tube. Another 30mL centrifuge tube was filled with tap water with the mass equal to the mass of 30mL centrifuge tube containing cellular sample. | ||
* | * The sample and water weight for balance were centrifuged in _______instrument with the following conditions: | ||
Speed: 18,000rpm | |||
Temperature: 4°C | |||
Time elapsed: 2 hours | |||
Rotor used: | |||
* After centrifugation, supernatants and pellet form. The pellet was discarded, and about 100mL supernatant yielded and was kept in a 500mL beaker. | |||
* The supernatant was filtered through Supor®-450 47mm membrane filter by pouring the supernatant through filter with membrane attached. Filter was attached to pressure suction that draws the supernatant through the membrane filter. The supernatant was filtered into a 800mL Erlenmeyer flask, then poured into a 25mL falcon tube. | |||
* The filtered protein supernatant was injected into the GE Pharmacia® AKTA Purifier 100 Fast Protein Liquid Chromatography to separate ADA proteins from the rest of proteins in sample through the following procedure: | |||
** | |||
==Result for Protein Extraction and Purification from BL21(DE3) Cells== | ==Result for Protein Extraction and Purification from BL21(DE3) Cells== | ||
Revision as of 10:09, 24 November 2012
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Purpose
Procedure for Protein Extraction and Purification from BL21(DE3) Cells
Speed: 18,000rpm Temperature: 4°C Time elapsed: 2 hours Rotor used:
Result for Protein Extraction and Purification from BL21(DE3) Cells
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