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University of Wisconsin–Madison

Mass Spectrometry / Proteomics Facility

We are open for business! We are happy to be back. During Phase 1, we will be operating at reduced capacity. We will make every effort to keep turnaround times short as we understand there are many projects that have been waiting. We appreciate your patience and business. The UW Biotechnology Center is following University guidelines regarding COVID-19.

Buffers and Solutions for In Gel Digests

500mM Ammonium Bicarbonate [(NH4)HCO3]

  • (NH4)HCO3-1.6g
  • Millipore water-to 40ml final volume
  • Filter through high protein binding filter [Nitrocelulose or PTFE; 0.45um pore size)

25 mM (NH4)HCO3

  • 500mM (NH4)HCO3-2ml
  • Millipore water-to 40ml final volume
  • Check pH (should be between 8-8.5)

25mM (NH4)HCO3/50% Acetonitrile [ACN]

  • 500mM (NH4)HCO3-2ml
  • ACN (HPLC grade)-20ml
  • Millipore water-18ml
  • Make sure pH stays at 8-8.5

500mM Ditiothreitol [DTT] stock

  • DTT-77.1mg
  • 25mM (NH4)HCO3-to 1ml final volume
  • Store at -20°C in 50ul aliquots

550mM Iodoacetamide [IAA] stock

  • IAA-101.7mg
  • 25mM (NH4)HCO3-to 1ml final volume
  • Store at -20°C in 50ul aliquots

30mM Potassium Ferricyanide (III) [K3Fe(CN)6]

  • K3Fe9(CN)6-494mg
  • Millipore water-to 50ml final volume
  • Wrap in aluminum foil and store at 4°C up to three months.

100mM Sodium Thiosulfate [Na2S2O3]

  • Na2S2O3-237mg
  • Millipore water-to 50ml final volume
  • Store at 4°C up to three months

Trypsin stock

  • Trypsin [PROMEGA Modified Sequence Grade]-20ug
  • 25mM (NH4)HCO3-1ml (resuspend gently at room temperature)
  • Store at -20°C in 100ul aliquots

10% N-octyl Glucoside

  • N-octyl Glucoside [SIGMA]-250mg
  • Millipore water-to 2.5ml final volume
  • Store at room temperature

0.1% ProteasMAX stock

  • ProteasMAX [Promega, Madison, WI]- 1mg
  • 25mM (NH4)HCO3-1ml
  • Store at -20°C in 50ul aliquots (single use only)
  • Dilute to 0.01% working concetration in a final digestion reaction