Protocols
System Suitability (CompRef) Study

Dataset: CompRef_Proteome_UNC_R1_QE

Analytical Sample Protocol

  1. Starting Amount 1 mg of P5 and P6 of crysopulvarized tissue (5-10 % total protein extracted)
  2. Proteolysis Trypsin
  3. Alkylation Iodoacetic acid
  4. Enrichment N/A
  5. Labelling Label-Free
  6. Fractionation Strong Cation Exchange (SCX)
  7. Fractions 24


Chromatography Protocol

  1. Column Type C18
  2. Column Length 45 cm
  3. Inside Diameter 75 µm
  4. Particle Type silica
  5. Particle Size 3.5 µm
  6. Injected 8 µL (~ 8 mg on-colµmn per LC-MS run)
  7. Gradient Length 240 min


Mass Spectrometry Protocol

  1. Type High resolution Orbitrap mass analysis
  2. Instrument Q Exactive
  3. Dissociation High-energy collision-induced dissociation
  4. MS 1 Resolution 70,000
  5. MS 2 Resolution 17,500
  6. Collision Energy NCE 27
  7. Precursors Top 20