SDS PAGE Sample Buffer
SB is
10 % by volume glycerol
62.5 mM TrisHCL, pH 6.8
2 % by weight SDS
0.01 mg/mL BfB
5% (Janet) or 2% (Red book) by volume bME to be added after thawing.
Stock solutions and materials
- 1 M Tris, pH 6.8
- Glycerol neat
- SDS solid
- bME neat
- Bfb slurry 0.5 % by mass (5 mg/ml)
To make 10 mL of 4X SB (less bME)
- 4 mL glycerol
- 0.8 g SDS
- 2.5 mL TrisáHCl, pH 6.8 stock
- 0.08 mL BfB slurry
- Add water to bring to 8 mL
- Divide into aliquots leaving room for 20% volume bME and store at -20 C. For
example make 10 0.8 mL aliquots.
- Thaw aliquots and add 20 % mL bME, for example 0.2 mL bME to 0.8 mL aliquots.
To make 2X add and equal volume of H2O to the 4X.
Reference Recipe for 100 mL 2X SB from Red Book
- 25 mL buffer A
- 20 mL glycerol
- 4 g SDS
- (2 mL bME) add to aliquots note this would be 2% bME, Janet uses 10%
- 1 mg BfB
- H2O to 100 mL
Buffer A = 4X TrisCl/SDS pH 6.8
0.5 M TrisCl and 0.4% SDS
To make 0.5 mL of 2X SB, Red Book version
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