User:Paul Rothenberg/Notebook/Pauls Notebook/2014/06/17
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Dialysis of Apo Mb
I set the Apo Mb prepared yesterday to run dialysis overnight. We will take a UV-VIS spectrum of it tomorrow.
Note -- it had to be moved from the cold room to the fridge in the lab because of cold room maintenance. Will check on it tomorrow.
Purification of CoMb
Andrew and I are going to purify the CoMb we have through the PD-10 column. We are running it with H2O in the same method we have previously.
After collecting excess porphyrin, we ran 25 mL of 1 M NaOH/NaCl to clear the column, and re-equilibrated with 25 mL of H2O before adding the next aliquot of protein.
Preparation of Mb Citrate Buffer
We are also going to prepare a standard Mb Citrate solution to lyophilize. For a 10 mM Citrate (pH 3) buffer,
We have 500 mg of KCl already measured out. We put 5 mg of Mb in, and added 20 mL of buffer.
All are 10 mM buffers