User:Paul Rothenberg/Notebook/Pauls Notebook/2014/06/09

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(Preparation of more Apo-Mb)
(Preparation of more Apo-Mb)
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==Preparation of more Apo-Mb==
==Preparation of more Apo-Mb==
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Andrew and I are going to synthesize more Apo-Mb for preparation of Co-Mb. We will follow the same protocol as I did [http://openwetware.org/wiki/User:Paul_Rothenberg/Notebook/Pauls_Notebook/2014/03/05 here]. We extracted about 7 mL of apoprotein. We also had to mix pH 2 H<sub>2</sub>O with equal parts 2-butanone because solid clumps blocked the separatory funnel. We added 4 mL of each for a total volume of 16 mL before separation. We added the apoprotein to a dialysis cassette and let run overnight.
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Andrew and I are going to synthesize more Apo-Mb for preparation of Co-Mb. We will follow the same protocol as I did [http://openwetware.org/wiki/User:Paul_Rothenberg/Notebook/Pauls_Notebook/2014/03/05 here]. We made a 0.05 M Tris solution, dissolved 200 mg of Mb into it, and added 40 μL of 1N HCl to bring the pH to 2.3. We then extracted with 4 mL of 2-butanone for two rounds.
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We extracted about 7 mL of apoprotein. We also had to mix pH 2 H<sub>2</sub>O with equal parts 2-butanone because solid clumps blocked the separatory funnel. We added 4 mL of each for a total volume of 16 mL before separation. We added the apoprotein to a dialysis cassette and let run overnight in a 10 mM Sodium Bicarbonate buffer.
==Synthesis of Co-Mb==
==Synthesis of Co-Mb==

Revision as of 10:55, 9 June 2014

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Preparation of more Apo-Mb

Andrew and I are going to synthesize more Apo-Mb for preparation of Co-Mb. We will follow the same protocol as I did here. We made a 0.05 M Tris solution, dissolved 200 mg of Mb into it, and added 40 μL of 1N HCl to bring the pH to 2.3. We then extracted with 4 mL of 2-butanone for two rounds.

We extracted about 7 mL of apoprotein. We also had to mix pH 2 H2O with equal parts 2-butanone because solid clumps blocked the separatory funnel. We added 4 mL of each for a total volume of 16 mL before separation. We added the apoprotein to a dialysis cassette and let run overnight in a 10 mM Sodium Bicarbonate buffer.

Synthesis of Co-Mb


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