ACIDIC-NATIVE GEL Protocol( For Basic Proteins (pI7.0))
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1953
USE ONLY FRESH GELS
Stock Solutions
1) 1.5M Acetate-KOH pH 4.3 (48ml 1MKOH + 17.2ml AcH + H2O up to 200ml) - Keep RT.
2) 30% Acrylamide 0.8% Methylene bis Acrylamide. Keep 4°C.
3) 0.25M Acetate-KOH pH 6.8 (48ml 1MKOH + 2.9ml AcH + H2O up to 200ml) - Keep RT.
4) 10% Ammonium Persulfate (APS). Keep 4°C less than 1 month.
Running Buffer x1
| <center> ß-Alanine</center> | <center> 0.35M</center> | <center> 18.7gr</center> |
| <center> AcH</center> | <center> 0.14M </center> | <center> 4.8ml </center> |
| <center> H2O up to</center> | <center> 600ml</center> | |
| <center> Adjust pH to</center> | <center> 4.3</center> |
Keep at RT
Dissolving Buffer x5
| <center> Glycerol 50%</center> | <center> 1.45ml</center> |
|
<center>
0.25M Acetate-KOH pH 6.8 </center>
|
<center> 0.5ml</center> |
|
<center>
Methyl Green</center>
|
<center>
traces</center>
|
Keep in aliquots of 1ml at -20C
