User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/03: Difference between revisions
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==Procedure== | ==Procedure== | ||
* Column from | * Column from purifier were washed with 100mL binding buffer at pH 7.5 with 500nM Tris, 200nM NaCl, and 30mM of imidazol. | ||
* 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. | ||
* For specific parameter for the column purifier, please refer to [[User:Melissa Novy/Notebook/CHEM-571/2012/10/03|Melissa's Notebook]] for the details of the column purifier and conditions which the ADA protein was run at. | |||
==Discussion== | ==Discussion== |
Revision as of 20:39, 26 October 2012
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Purpose
Procedure
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