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STS 299 |
2-DE Protein Separation
The Geneva Proteomics Core Facility uses a sigmoidal immobilized pH gradient (IPG) for
isoelectric focusing and the usual vertical SDS polyacrylamide gel with
some small modifications (PDA as crosslinker and sodium thiosulfate as
adjunct catalyst) for the second dimension. A sensitive ammoniacal silver
staining is used as a non-radioactive protein detection method. The
resulting 2-D PAGE is scanned on a laser densitometer. The digital image may
then be retrieved from the ExPASy server and analysed at your site using,
for example, the ImageMaster 2D Platinum analysis software. Alternatively, a paper
print may be sent by mail (Disclaimer).
Sample preparation
Sample preparation and solubilization are crucial factors for the overall
performance of the 2-D PAGE technique. Protein complexes and aggregates
should be completely broken up in order to avoid the appearance of new spots
due to a partial protein solubilization. Thus, samples have to be sent to
the Geneva Proteomics Core Facility either in a fresh solution containing 8 M urea, 4% (w/v)
CHAPS, Tris base 40 mM, DTE 65 mM and a trace of bromophenol blue (see below).
- Minigels
Bromophenol blue (in a concentration between 1 and 5 mg/ml), or as
dried powder. The minimum amount of protein has to be around
25 µg.
The sample may contain salt in amounts lower than 100 mM.
- Maxi gels
Bromophenol blue (in a concentration between 2 and 10 mg/ml), or as
dried powder. The minimum amount of protein has to be around
100 µg.
The sample may contain salt in amounts lower than 100 mM.
Put your samples in an Eppendorf tube, freeze them at -20 degree Celsius
and put them into dry ice for shipment.
Prices
Method | Swiss Fr. / sample 1-10 samples | Swiss Fr. / sample > 10 samples |
Minigels 2-DE and Silver staining (8 x 9 cm) | 125.00 | Inquire |
Minigels 2-DE and Coomassie staining | 100.00 | Inquire |
Minigels 2-DE and PVDF transfert | 150.00 | Inquire |
Maxi 2-DE and Silver staining (18 x 20 cm) | 250.00 | Inquire |
Maxi 2-DE and Coomassie staining | 200.00 | Inquire |
Maxi 2-DE and PVDF transfert | 300.00 | Inquire |
Indicated prices takes into account that you are coming from Faculty of Medecine, Uni Geneve or HUG
Minigels 2-DE and Silver staining (8 x 9 cm) | 200.00 | Inquire |
Minigels 2-DE and Coomassie staining | 175.00 | Inquire |
Minigels 2-DE and PVDF transfert | 250.00 | Inquire |
Maxi 2-DE and Silver staining (18 x 20 cm) | 450.00 | Inquire |
Maxi 2-DE and Coomassie staining | 400.00 | Inquire |
Maxi 2-DE and PVDF transfert | 500.00 | Inquire |
Prices do not include VAT (7.6%).
Time delay
Allow two weeks for delivery of the results (prints and images).