User:Matt Hartings/Notebook/Photosynthesis/2014/12/09: Difference between revisions

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## 0.225mL Mb solution added after 3 scans  
## 0.225mL Mb solution added after 3 scans  
## Should see this in the spectra
## Should see this in the spectra
*Note
# By the end of the day, the 30% peroxide that I had taken out of the fridge was showing bubbles. Concentration of peroxide should still be greater than that of OPD.
# Both the Mb in isobutanol and the KCl in isobutanol gave reddish solutions of roughly the same intensity at the end of the reaction. I will have to revisit these. Perhaps I'll do that tomorrow.


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Revision as of 13:11, 9 December 2014

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Objective

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Samples

Sample 1

Mb lyophilized from Tris and KCl in water. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg myoglobin from Tris KCl
    2. OPD
      1. 2.6mg OPD in 2.6mL water
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL water
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: Mb_OPD_H2O2_tris_water_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 2

lyophilized from Tris and KCl in water. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin - no myoglobin this run
      1. 15mg lyophilized Tris KCl
    2. OPD
      1. 2.5mg OPD in 2.5mL water
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL water
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: KCl_OPD_H2O2_tris_water_25_SCANNUMBER.txt
    6. 0.225mL KCl solution added after 3 scans
    7. Should see this in the spectra

Sample 3

Mb lyophilized from Tris and KCl in methanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg myoglobin from Tris KCl in 1mL 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 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: Mb_OPD_H2O2_tris_meoh_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 4

lyophilized Tris and KCl in methanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg Tris KCl in 1mL methanol
    2. OPD
      1. 2.5mg OPD in 2.5mL methanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL methanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: KCl_OPD_H2O2_tris_meoh_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 5

Mb lyophilized from Tris and KCl in ethanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg myoglobin from Tris KCl in 1mL ethanol
    2. OPD
      1. 3.7mg OPD in 3.7mL ethanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL ethanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: Mb_OPD_H2O2_tris_etoh_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 6

lyophilized Tris and KCl in ethanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg Tris KCl in 1mL ethanol
    2. OPD
      1. 3.4mg OPD in 3.4mL ethanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL ethanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: KCl_OPD_H2O2_tris_etoh_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 7

Mb lyophilized from Tris and KCl in isopropanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg myoglobin from Tris KCl in 1mL isopropanol
    2. OPD
      1. 3.0mg OPD in 3.0mL isopropanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL isopropanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: Mb_OPD_H2O2_tris_isop_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 8

lyophilized Tris and KCl in isopropanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg Tris KCl in 1mL isopropanol
    2. OPD
      1. 2.5mg OPD in 2.5mL isopropanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL isopropanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: KCl_OPD_H2O2_tris_isop_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 9

Mb lyophilized from Tris and KCl in 1-propanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg myoglobin from Tris KCl in 1mL 1-propanol
    2. OPD
      1. 2.9mg OPD in 2.9mL 1-propanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL 1-propanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: Mb_OPD_H2O2_tris_1prop_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 10

lyophilized Tris and KCl in 1-propanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg Tris KCl in 1mL 1-propanol
    2. OPD
      1. 4.0mg OPD in 4.0mL 1-propanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL 1-propanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: KCl_OPD_H2O2_tris_1prop_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 11

Mb lyophilized from Tris and KCl in 1-butanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg myoglobin from Tris KCl in 1mL 1-butanol
    2. OPD
      1. 2.7mg OPD in 2.7mL 1-butanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL 1-butanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: Mb_OPD_H2O2_tris_1but_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 12

lyophilized Tris and KCl in 1-butanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg Tris KCl in 1mL 1-butanol
    2. OPD
      1. 5.0mg OPD in 5.0mL 1-butanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL 1-butpanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: KCl_OPD_H2O2_tris_1but_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 13

Mb lyophilized from Tris and KCl in isobutanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg myoglobin from Tris KCl in 1mL isobutanol
    2. OPD
      1. 3.0mg OPD in 3.0mL isobutanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL isobutanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: Mb_OPD_H2O2_tris_isob_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra

Sample 14

lyophilized Tris and KCl in isobutanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans

  1. Stock solutions:
    1. Myoglobin
      1. 15mg Tris KCl in 1mL isobutanol
    2. OPD
      1. 3.0mg OPD in 3.0mL isobutanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL isobutanol
  2. Experimental Run
    1. Sample chamber set to 25C 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. 200 scan average
      4. Perform a scan every 10 seconds
      5. Stop after 10minutes
    5. Files saved as: KCl_OPD_H2O2_tris_isob_25_SCANNUMBER.txt
    6. 0.225mL Mb solution added after 3 scans
    7. Should see this in the spectra


  • Note
  1. By the end of the day, the 30% peroxide that I had taken out of the fridge was showing bubbles. Concentration of peroxide should still be greater than that of OPD.
  2. Both the Mb in isobutanol and the KCl in isobutanol gave reddish solutions of roughly the same intensity at the end of the reaction. I will have to revisit these. Perhaps I'll do that tomorrow.


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