User:Matt Hartings/Notebook/Photosynthesis/2013/04/03

From OpenWetWare

Jump to: navigation, search
Protein Re-engineering Main project page
Previous entry      Next entry



Mb catalysis DMSO

Following the description from Tuesday. Stock solutions used

  1. 28.6mg of o-phenylenediamine in 26.5mL of DMSO
  2. 1.134 ml 30% H2O2 +8.866 ml DMSO
  3. 11.1mg of (5.7mg myoglobin +0.4861g KCl lyophilized from 20mL of 10mM citrate pH 3) in 1 mL of DMSO.

Samples were made by

  1. 2.025 mL of the OPD solution
  2. 0.225 mL of the Mb solution
  3. Start the scan
    1. 1,000 us integration time
    2. Average over 500 scans
    3. Run for 10 minutes (600,000,000 us)
  4. Add 0.75mL of the H2O2 solution
    1. Added after scan reached 1%
  5. Kinetics scans were run at 25, 35, and 45C (DMSO freezes at around 20C). Several before and after spectra were also taken, as were the dark and reference signal spectra.
  6. Absorbance measurements were calculated using the following equation: A = -log[(SampleSignal-DarkSignal)/(ReferenceSignal-DarkSignal)]

Mb Catalysis Isopropanol

Following the description from Tuesday. Stock solutions used

  1. 29.2mg of o-phenylenediamine in 27mL of Isopropanol
  2. 1.134 ml 30% H2O2 +8.866 ml Isopropanol
  3. 15mg of (5.7mg myoglobin +0.4861g KCl lyophilized from 20mL of 10mM citrate pH 3) in 1.5 mL of isopropanol.

Samples were made by

  1. 2.025 mL of the OPD solution
  2. 0.225 mL of the Mb solution
  3. Start the scan
    1. 1,000 us integration time
    2. Average over 500 scans
    3. Run for 10 minutes (600,000,000 us)
  4. Add 0.75mL of the H2O2 solution
    1. Added after scan reached 1%
  5. Kinetics scans were run at 5, 15, 25, 35, and 45C. Several before and after spectra were also taken, as were the dark and reference signal spectra.
  6. Absorbance measurements were calculated using the following equation: A = -log[(SampleSignal-DarkSignal)/(ReferenceSignal-DarkSignal)]


All of the kinetics data taken today are no good because I didn't save the dark and reference spectra as Signal files. They were saved as processed absorption spectra. DMSO showed reactivity while there was no reactivity in isopropanol.


Personal tools