User:Tamra L. Fisher/Notebook/Research for Dr. Hartings/2012/07/16: Difference between revisions
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==Reconstitute Asc Hb with Nickel== | ==Reconstitute Asc Hb with Nickel== | ||
* After the Asc Hb with Nickel sat undisturbed overnight, the protein was poured into centrifuge tubes and spun at 18000rpm for two hours at 4°C. | * After the Asc Hb with Nickel sat undisturbed overnight, the protein was poured into centrifuge tubes and spun at 18000rpm for two hours at 4°C. | ||
* The protein was then purified over a PD-10 Desalting column: | |||
# A PD-10 Desalting column was prepared with one column volume of dH<sub>2</sub>O was run over the column, and two column volumes of 25mM Tris, pH 8.3 run over the column. | |||
# 2.5mL of the dialyzed Reconstituted Asc Hb was then run over the column. | |||
# The protein was eluted off with 3.5mL of 25mM Tris, pH 8.3. | |||
# The loading and elution of the protein was repeated many times. | |||
==Purifying the Asc Hb with Manganese== | ==Purifying the Asc Hb with Manganese== |
Revision as of 13:57, 16 July 2012
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Reconstitute Asc Hb with Nickel
Purifying the Asc Hb with Manganese
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