Objective
Take kinetic data for Mb (lyophilized from KCl/phosphate buffer) catalyzed o-phenylenediamine + H2O2 reaction in water (45C) and Mb (lyophilized from KCl/tris buffer) catalyzed o-phenylenediamine + H2O2 reaction in water (35, 45C).
Procedure and Data
sample 1
- Stock solutions:
- Myoglobin
- 1.4mg (Mb:KCl lyophilized from Phosphate, 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 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_phosphate_water_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
sample 1
- Stock solutions:
- Myoglobin
- 1.2mg (Mb:KCl lyophilized from Tris, 5mg:500mg) in 1.0mL water
- OPD
- 3.0mg OPD in 3.0mL water
- H2O2
- 0.114mL 30% H2O2 in 0.887mL water
- 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_tris_water_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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