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

From OpenWetWare
Jump to navigationJump to search
Error creating thumbnail: Unable to save thumbnail to destination Protein Re-engineering Main project page
Previous entry      Next entry



Objective

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

Procedure and Data

sample 1

  1. Stock solutions:
    1. Myoglobin
      1. 1.3mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL water
    2. OPD
      1. 3.5mg OPD in 3.5mL water
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL water
  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_water_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. 2.5mg OPD in 2.5mL water
    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_Acetate_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. same as above
    2. OPD
      1. 3.1mg OPD in 3.1mL water
    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_Acetate_meoh_15_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



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Matt_Hartings/Notebook/Photosynthesis/2013/10/10&oldid=1013484"