Team:EPF-Lausanne/Protocol/Silver staining

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For a 5% Stacking gel,
{| class="wikitable" style="text-align: center; color: black;"
{| class="wikitable" style="text-align: center; color: black;"
|'''Stacking gel'''
|'''Stacking gel'''
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|'''Volume, en mL'''
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|Gel percentage
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|5 %
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|30% Polyacrylamide
|30% Polyacrylamide
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|1.36
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|1M Tris (pH 6.8)
|1M Tris (pH 6.8)
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|10% Ammonium persulfate
|10% Ammonium persulfate
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|0.08 mL
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|0.08
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|10% SDS
|10% SDS
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|0.08 mL
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|0.08
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|TEMED
|TEMED
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|0.008 mL
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|0.008
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|H2O
|H2O
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|5.44 mL
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|5.44
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|Total volume
|Total volume

Revision as of 00:00, 27 September 2012

Protocol: Silver Staining


Preparation of Gel

Ingredients

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


Team-EPF-Lausanne ssp.jpg




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