User:Matt Hartings/Notebook/Photosynthesis/2013/10/17

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Objective

Take kinetic data for Mb (lyophilized from KCl/acetate buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (45C). Take kinetic data for Mb (lyophilized from KCl/phosphate buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (5C, 15C, and 25C).

Procedure and Data

sample 1

  1. Stock solutions:
    1. Myoglobin
      1. 1.3mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL methanol
    2. OPD
      1. 3.6mg OPD in 3.6mL methanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL methanol
  2. Experimental Run
    1. Sample chamber set to 45C and to stir
    2. N2 run through the cuvette holder to reduce condensation on cuvette
    3. 2.025mL OPD stock solution
    4. 0.75mL H2O2 stock solution
      1. Start Data Collection
      2. 1ms Integration
      3. 10 scan average
      4. Wait 1 minute after each scan
      5. Stop after 2 hours
    5. Files saved as: Mb_OPD_H2O2_Acetate_meoh_45_SCANNUMBER.txt
    6. 0.225mL Mb stock solution added just before the third scan
    7. This is denoted in the data file for the third scan

sample 2

  1. Stock solutions:
    1. Myoglobin
      1. 1.4mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL methanol
    2. OPD
      1. 2.4mg OPD in 2.4mL methanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL methanol
  2. Experimental Run
    1. Sample chamber set to 5C and to stir
    2. N2 run through the cuvette holder to reduce condensation on cuvette
    3. 2.025mL OPD stock solution
    4. 0.75mL H2O2 stock solution
      1. Start Data Collection
      2. 1ms Integration
      3. 10 scan average
      4. Wait 1 minute after each scan
      5. Stop after 2 hours
    5. Files saved as: Mb_OPD_H2O2_phosphate_meoh_5_SCANNUMBER.txt
    6. 0.225mL Mb stock solution added just before the third scan
    7. This is denoted in the data file for the third scan

sample 3

  1. Stock solutions:
    1. Myoglobin
      1. No myoglobin was used in this sample
    2. OPD
      1. 3.6mg OPD in 3.6mL methanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL methanol
  2. Experimental Run
    1. Sample chamber set to 5C and to stir
    2. N2 run through the cuvette holder to reduce condensation on cuvette
    3. 2.025mL OPD stock solution
    4. 0.75mL H2O2 stock solution
      1. Start Data Collection
      2. 1ms Integration
      3. 10 scan average
      4. Wait 1 minute after each scan
      5. Stop after 2 hours
    5. Files saved as: OPD_H2O2_phosphate_meoh_5_SCANNUMBER.txt
    6. 0.225mL Mb stock solution added just before the third scan
    7. This is denoted in the data file for the third scan



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