User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/03: Difference between revisions
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==Procedure== | ==Procedure== | ||
* Column from AKTA purifier were washed with 100mL binding buffer at pH 7.5 with Tris, NaCl, and 30mM of imidazol. | * Column from AKTA purifier were washed with 100mL binding buffer at pH 7.5 with Tris, NaCl, and 30mM of imidazol. | ||
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 17:56, 7 October 2012 (EDT)''':what are the tris nad NaCl concnetrations? what is AKTA purifier? | |||
* Protein supernatant filtered from yesterday was injected into column. | * Protein supernatant filtered from yesterday was injected into column. | ||
* Protein supernatant were run through the column with AKTA purifier. Flow through were collected in 50ml falcon tube just in case protein failed to bind to column. | * Protein supernatant were run through the column with AKTA purifier. Flow through were collected in 50ml falcon tube just in case protein failed to bind to column. | ||
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* UV-vis showed concentration protein elution at 280nm absorptivity, and indicated the elution test tube with most concentrated ADA protein. | * UV-vis showed concentration protein elution at 280nm absorptivity, and indicated the elution test tube with most concentrated ADA protein. | ||
* Two test tubes of ADA protein were collected in a 15mL falcon tube, and flow through of the protein are stored in 4C. | * Two test tubes of ADA protein were collected in a 15mL falcon tube, and flow through of the protein are stored in 4C. | ||
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 17:56, 7 October 2012 (EDT)''':more information - flow rates? time? any specific parameters for the FPLC. | |||
==Discussion== | ==Discussion== |
Revision as of 14:56, 7 October 2012
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