User:Matt Hartings/Notebook/Photosynthesis/2013/11/05

Protein Re-engineering

Main project page Previous entry      Next entry

Objective Take kinetic data for Mb (lyophilized from KCl/tris buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (25, and 35C). Procedure and Data sample 1 Stock solutions: Myoglobin 1.1mg (Mb:KCl lyophilized from Tris, 5mg:500mg) in 1.0mL methanol OPD 3.0mg OPD in 3.0mL methanol H2O2 0.114mL 30% H2O2 in 0.887mL methanol 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_meoh_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 2.5mg OPD in 2.5mL methanol H2O2 0.114mL 30% H2O2 in 0.887mL methanol Experimental Run Sample chamber set to 35C 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_meoh_35_SCANNUMBER.txt 0.225mL Mb stock solution added just before the third scan This is denoted in the data file for the third scan