LaCava Research Wiki

Initiated September 2017

%Heavy labeling

jlacava10th January 2020 at 11:01am

Cells grown in Lys8, Arg10

  • 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
    • Labeling efficiency should be ~95%, certainly better than 90% for using in an experiment
  • secondary check on proteins –> using MQ proteingroups.txt
    • good labeling is sufficient for mix after purification (MAP) SiLAC experiments
    • this factor is used to determine the expected %H of a 50:50 (by protein quantity) mixture of heavy and light materials used in IDiRT (or mix before purification SiLAC, which we don't really do)

Prior art on %H labeling > Taylor et al. Cell 2013