Objective
Take kinetic data for Mb catalyzed o-phenylenediamine + H2O2 reaction at higher temperatures (25, 35, and 45C)
Procedure and Data
Stock solutions
sample 1
- Stock solutions:
- Myoglobin
- 2.0mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL water
- OPD
- 4.1mg OPD in 2.4mL 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_Citrate_H20_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
- Note - This run was over in a hurry. Something was off (mass of Mb or mass of OPD or both) I stopped the run early and will use the balance in 207 for the remainder of experiments.
sample 2
- Stock solutions:
- Myoglobin
- 1.5mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL water
- OPD
- 3.0mg OPD in 3.3mL water
- H2O2
- 0.114mL 30% H2O2 in 0.887mL water
- 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_Citrate_H20_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
sample 3
- Stock solutions:
- Myoglobin
- Used the same stock as in sample 2
- OPD
- H2O2
- Experimental Run
- Sample chamber set to 45C 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_Citrate_H20_45_SCANNUMBER.txt
- 0.225mL Mb stock solution added just before the third scan
- This is denoted in the data file for the third scan
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