LaCava Research Wiki

Initiated September 2017

%Heavy mixing

jlacava10th January 2020 at 11:01am

Cells grown in Lys8, Arg10 mixed with those grown in natural Lys and Arg

  • Initial check on peptides –> using MQ peptides.txt
    • considering only 0 missed cleavages to make the calculation straight forward (assuming enough peptide for sufficient statistical power - what is the minimum number of peptides appropriate?)
    • these values are derived from the Ratio H/L data in peptides.txt
    • we convert this to %H by taking (ratio H/L / 1+ratio H/L)*100
  • secondary check on proteins –> using MQ proteingroups.txt
    • Mixing observed should be at the level predicted by the %H labeling AND 50:50 proportions between H and L materials
      • i.e. assuming calculated ~95% labeling efficiency, ~48% H in the mix would be obtained.
      • we like a threshhold of an actual value being within ~10% the expected value –> ~43-53% in the above example.
      • Otherwise, we can adjust the mix by adding more H or L powder
      • In any case, the deviation between expected and actual should be used to normalize results.