Geneva Proteomics Core Facility

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)	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).

2-DE Protein Separation

Sample preparation

Prices

Time delay

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Last modified 14/Nov/2007 by CHH