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Obtain proteins from cells prepared on
Adenosine deaminase (ADA) will be separated from the rest of the cell proteins on
The cells were lysed using a Misonix XL-2000 sonicator to allow direct manipulation of organelles, proteins, and other cell components in solution.
The sonicated cell solution was centrifuged at 4°C and 18,000 rpm for a period of 2 hours.
Note that cooling the solutions prevents cell proteins from denaturing.
The resultant supernatant was then filtered with a 450 nm nylon filter to remove all large cell components. The filtrate is expected to contain cell proteins.
The filtrate containing cell proteins, including ADA, was refrigerated at 4°C until needed.
Please refer to
Dhea Patel's entry
for the detailed protocol for protein purification and proper disposal of biowaste.
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