Team:EPF-Lausanne/Protocol/Silver staining

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(Silver Staining)
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==Silver Staining==
==Silver Staining==
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[[File:Team-EPF-Lausanne_ssp.jpg|600px]]
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[[File:Team-EPF-Lausanne_ssp.jpg|650px]]

Latest revision as of 00:07, 27 September 2012

Contents

Protocol: Silver Staining


Preparation of Gel

Note: Please, adapt the final percentage of polyacrilamide according to the size of the protein.


Expected protein size Final polyacrilamide concentration
4-40 kDA 20%
12-45 kDA 15%
10-70 kDA 12.5%
15-100 kDA 10%
25-200 kDA 8%

For a Gel percentage of 7.5 % the proportions are the following:

For a 5% Stacking gel,
Separating gel Volume, en mL
30% Polyacrylamide 10
1.5M Tris (pH 8.8) 10
10% Ammonium persulfate 0.4
10% SDS 0.4
TEMED 0.038
H2O 19.2
Total volume 40
Stacking gel Volume, en mL
30% Polyacrylamide 1.36
1M Tris (pH 6.8) 1 mL
10% Ammonium persulfate 0.08
10% SDS 0.08
TEMED 0.008
H2O 5.44
Total volume 8 mL


Gel Running

For running and transfering the gel, please refer to Wester Blot protocol.

Silver Staining

Team-EPF-Lausanne ssp.jpg