Please go through the CHECKLIST below !
NOTE 1 : failure to follow these guidelines may lead to poor results.
A typical example is unwashed cells, which may be heavily contaminated with albumin from culture media. This dramatically reduces proteome coverage.
NOTE 2 : only samples in 1.5 ml Eppendorf tubes are accepted. Other tube formats only after special agreements.
NOTE 3 : Tubes MUST be numbered sequentially : 1, 2, 3,…
1. Discussion
Preliminary discussion BEFORE sample preparation always helps and increases chances of sucess. It is mandatory for complex and large projects.
Pilot experiments are crucial to establish feasibility and optimize protocols. They can be done quickly and are not too expensive.
Group leaders : contact us during grant preparation if your project includes key proteomics experiments. We can provide advice, a letter of support and budget info.
2. Sample preparation: if you are submitting…
Cell pellets
- wash them 2x with excess buffer (PBS)
- spin, remove maximum of liquid
- freeze at -20C or -80C
- indicate on submission form :
- type of cells
- approx. number
Protein solutions
Indicate on submission form :
- full, exact composition of the buffer/solvent. We need to know about detergents (even traces), salts, pH
- an estimate of protein concentration and how it was measured
Affinity beads
- wash 1x-2x with excess PBS or Tris buffer, remove liquid
- freeze at -20C
- indicate on submission form :
- approx. volume of pelleted beads AND residual liquid
- if residual liquid is present, what is it ?
- type of beads : agarose, magnetic,… (ideally product # ?)
- expected amount and type of known protein present on the beads (Antibody, GFP-trap, …)
Tissues
- wash tissue 1x with cold PBS or Tris buffer, remove liquid
- weigh tissue and freeze at -80C
- indicate on submission form the weight of tissue piece
Polyacrylamide gel bands
- stain gel with MS-compatible Coomassie blue or Silver
- cut bands with a clean scalpel under laminar flow
- transfer band(s) to an Eppendorff tube, freeze -20C
- indicate on submission form
- approx MW on gel or
- attach image of gel with MW marker
3. Sample submission and delivery
- Label the tubes clearly with NUMBERS (1,2,3,…N)
- Store at -20C / -80C
- Fill-in the on-line submission form (see below). Please attach any relevant files
- Come and drop/ship the samples to the facility, best if under dry ice. Please contact us in advance when you are coming
All clear ? you can proceed to online sample submission