User:Paul Rothenberg/Notebook/Pauls Notebook/2013/07/15

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Reaction of OPD+H2O2 in Isopropanol Suspension with Mb/Tris Catalyst

We finished up the final trials of the reactions, and set the control buffers to lyopholize. They were flash frozen using the acetone-dry ice bath at -78°C.

Graphs:

Molar Absorptivity of 2,3 Diaminophenazine

We are now going to determine the molar absorptivity of 2,3 Diaminophenazine (DAPZ, to abbreviate it here).

Procedure

We started by making a 1mM solution in H2</subO, using 21.023 mg of 2,3 Diaminophenazine. It did not all dissolve in solution, so we added that to a 250 mL volumetric flask and added an additional 150 mL of water, making sure to dissolve any remaining solid in the previous flask.


This was also too strong, so we added the 250 mL to a 500 mL flask and added an additional 250 mL of water.


We remeasured out the DAPZ for a 50 μM concentration. It required 10.0 mg of DAPZ in 1 L of water. We also heated and stirred this to help any undissolved DAPZ dissolve in solution.

We then tried a 25 μM concentration by diluting the 50 μM solution with 1000 mL of water (dilution by a factor of 2). That was made, but we did not use that at all.


Dr. Hartings also suggested we dissolve the DAPZ in 1 mL of DMSO, and then add it to water. The concentration of this was 47,567 μM. Dissolving this in water, we got a 475.67 μM stock concentration.


We used this to make a 19, 28, 38, 48, and 57 μM solutions by adding 0.5 mL increments to the equivalent amounts of water for a total of 25 mL of solution.


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