User:Paul Rothenberg/Notebook/Pauls Notebook/2014/06/18: Difference between revisions
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Procedure for Today: | ===Procedure for Today:=== | ||
To synthesize more apoprotein, we made a stock solution of 50 mM Tris buffer at pH 7.2. We dissolved 50 mg of myoglobin into 20 mL of buffer, and placed it on ice. The pH was brought to 2.5 at 0°C with the addition of 3.2 mL of 1 N HCl. | To synthesize more apoprotein, we made a stock solution of 50 mM Tris buffer at pH 7.2. We dissolved 50 mg of myoglobin into 20 mL of buffer, and placed it on ice. The pH was brought to 2.5 at 0°C with the addition of 3.2 mL of 1 N HCl. Once the HCl was added, the solution was immediately mixed with equal volume of 2-butanone, shaken for 30 seconds, and let sit for 1 minute. | ||
Revision as of 07:32, 18 June 2014
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Reconstitution of ApoproteinGoing to reconstitute the Apoprotein with MnPPIX. Hopefully the yield will be greater today. Before that, I ran a UV-VIS spec of the apoprotein at 1/20 and pure to compare concentration values. It looks like we are losing product during dialysis.
Procedure for Today:To synthesize more apoprotein, we made a stock solution of 50 mM Tris buffer at pH 7.2. We dissolved 50 mg of myoglobin into 20 mL of buffer, and placed it on ice. The pH was brought to 2.5 at 0°C with the addition of 3.2 mL of 1 N HCl. Once the HCl was added, the solution was immediately mixed with equal volume of 2-butanone, shaken for 30 seconds, and let sit for 1 minute.
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