User:Matt Hartings/Notebook/Photosynthesis/2013/10/29

Protein Re-engineering

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Objective Take kinetic data for Mb (lyophilized from KCl/tris buffer) catalyzed o-phenylenediamine + H2O2 reaction in water (25, 15, and 5C). Procedure and Data sample 1 Stock solutions: Myoglobin 1.3mg (Mb:KCl lyophilized from Tris, 5mg:500mg) in 1.0mL water OPD 2.3mg OPD in 2.3mL water H2O2 0.114mL 30% H2O2 in 0.887mL water Experimental Run Sample chamber set to 25C and to stir N2 run through the cuvette holder to reduce condensation on cuvette 2.025mL OPD stock solution 0.75mL H2O2 stock solution Start Data Collection 1ms Integration 10 scan average Wait 1 minute after each scan Stop after 2 hours Files saved as: Mb_OPD_H2O2_tris_h2o_25_SCANNUMBER.txt 0.225mL Mb stock solution added just before the third scan This is denoted in the data file for the third scan sample 2 Stock solutions: Myoglobin same as above OPD 2.6mg OPD in 2.6mL water H2O2 0.114mL 30% H2O2 in 0.887mL water Experimental Run Sample chamber set to 15C and to stir N2 run through the cuvette holder to reduce condensation on cuvette 2.025mL OPD stock solution 0.75mL H2O2 stock solution Start Data Collection 1ms Integration 10 scan average Wait 1 minute after each scan Stop after 2 hours Files saved as: Mb_OPD_H2O2_tris_h2o_15_SCANNUMBER.txt 0.225mL Mb stock solution added just before the third scan This is denoted in the data file for the third scan