SDS-polyacrylamide gel electrophoresis

SOLUTIONS

30% acrylamide mix (30:0.8 bis)	30% (w/v) acrylamide
	0.8% (w/v) N,N'- methylenebisacrylamide
5x Electrophoresis buffer	Tris base: 15.1 g
	Glycice: 94 g
	SDS: 5 g
	H2O upto1 liter
4x Resolving-gel buffer	1.5M Tris-Cl (pH 8.8)
8x Stacking-gel buffer	1M Tris-Cl (pH 6.8)
1x SDS gel-loading buffer	50mM Tris-Cl (pH 6.8)
	100mM dithiothreitol
	2% SDS
	0.1% bromophenol blue
	10% glycerol

RESOLVING GEL MIXTURE

Resolving Gel (30ml)	6%	8%	10%	12%	15%
H2O	16.2	14.2	12.2	10.2	7.2
30% acrylamide mix	6.0	8.0	10	12	15
4x resolving-gel buffer	7.5	7.5	7.5	7.5	7.5
20% SDS	0.15	0.15	0.15	0.15	0.15
20% APS	0.15	0.15	0.15	0.15	0.15
TEMED	0.024	0.018	0.012	0.012	0.012

Resolving Gel (10ml)	6%	8%	10%	12%	15%
H2O	5.4	4.7	4.1	3.4	2.4
30% acrylamide mix	2.0	2.7	3.3	4	5
4x resolving-gel buffer	2.5	2.5	2.5	2.5	2.5
20% SDS	0.05	0.05	0.05	0.05	0.05
20% APS	0.05	0.05	0.05	0.05	0.05
TEMED	0.008	0.006	0.004	0.004	0.004

STACKING GEL MIXTURE

Stacking Gel (10ml)
	H2O	6.9 ml
	30% acrylamide mix	1.7 ml
	8x stacking-gel buffer	1.25 ml
	20% SDS	0.05 ml
	20% APS	0.05 ml
	TEMED	0.01 ml

SOLUTIONS FOR STAINING AND DESTAINING

Staining solution	methanol	90 ml
	H2O	90 ml
	acetic acid	20 ml
	Coomassie Brilliant Blue R250	0.25 g
	Dissolve well and filtrate through a filter paper.
Destaining solution	isopropanol	12.5%
	glacial acetic acid	10%
	Addition of small pieces of used X-ray film with the gel will accelerate the destaining process.

SOLUTION FOR DRYING

20% methanol, 3% glycerol in diltilled water

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