PAGE-SDS Laemmli Protocol
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Stock Solutions
1) 1.5M TrisHCl pH 8.3 + 0.4% SDS (adjust pH before you add the SDS). Keep RT.2) 30% Acrylamide 0.8% Methylene bis Acrylamide or 40% Acrylamide / Methylene bis Acrylamide (ratio: 37.5:1). Keep 4°C
3) 0.5M TrisHCl pH 6.8+ 0.4% SDS (adjust pH before you add the SDS). Keep RT.
4) 10% Ammonium Persulfate (APS). Keep 4°C less than 1 month.
Running Buffer x5
| <center> Trizma</center> | <center> 0.125M</center> | <center> 15.1gr</center> |
| <center> Glycine</center> | <center> 0.96M </center> | <center> 72.0gr </center> |
| <center> SDS</center> | <center> 0.5%</center> | <center> 5.0gr</center> |
| <center> H2O up to</center> | <center> 1 liter</center> |
Keep at RT
Dissolving (Sample) Buffer x5
| <center> Glycerol</center> | <center> 5ml</center> |
| <center> SDS</center> | <center> 1gr</center> |
| <center> ß Mercapto Ethanol<br /> (or 0.25M DTT)</center> | <center> 2.56ml</center> |
|
<center>
0.5M TrisHCl pH 6.8+ 0.4% SDS </center>
|
<center> 2.13ml</center> |
|
<center>
Bromo Phenol Blue</center>
|
<center>
traces</center>
|
