SDS-PAGE

From Genome@Yale

RESOLVING - 10mL					RESOLVING - 20mL				
	dH2O	30%	1.5M	10% SDS				dH2O	0.3	1.5M	10% SDS
4%	6.1	1.3	2.5	0.1			4%	12.2	2.6	5	0.2
5%	5.7	1.7	2.5	0.1			5%	11.4	3.4	5	0.2
6%	5.4	2	2.5	0.1			6%	10.8	4	5	0.2
7%	5.1	2.3	2.5	0.1			7%	10.2	4.6	5	0.2
8%	4.7	2.7	2.5	0.1			8%	9.4	5.4	5	0.2
9%	4.4	3	2.5	0.1			9%	8.8	6	5	0.2
10%	4.1	3.3	2.5	0.1			10%	8.2	6.6	5	0.2
11%	3.7	3.7	2.5	0.1			11%	7.4	7.4	5	0.2
12%	3.4	4	2.5	0.1			12%	6.8	8	5	0.2
13%	3.1	4.3	2.5	0.1			13%	6.2	8.6	5	0.2
14%	2.7	4.7	2.5	0.1			14%	5.4	9.4	5	0.2
15%	2.4	5	2.5	0.1			15%	4.8	10	5	0.2
16%	2.1	5.3	2.5	0.1			16%	4.2	10.6	5	0.2
17%	1.7	5.7	2.5	0.1			17%	3.4	11.4	5	0.2
18%	1.4	6	2.5	0.1			18%	2.8	12	5	0.2
19%	1.1	6.3	2.5	0.1			19%	2.2	12.6	5	0.2
20%	0.7	6.7	2.5	0.1			20%	1.4	13.4	5	0.2
10uL TEMED						20uL TEMED				
50uL 10% APS						100uL 10% APS

STACKING - 5mL							STACKING - 10mL				
	dH2O	30%	0.5M	10% SDS					dH2O	30%	0.5M	10% SDS
4%	3.05	0.65	1.25	0.05				4%	6.1	1.3	2.5	0.1
10uL TEMED							10uL TEMED
25uL 10% APS							25uL 10% APS

2X Laemmli Sample Buffer
                       5mL     50mL
0.5M Tris, pH 6.8      1mL     10mL
10% SDS                2mL     20mL
80% glycerol           1.25mL  12.5mL
beta-mercaptoethanol   0.5mL   5mL
5% bromophenol blue    0.1mL   1mL
dH2O                   0.15mL  1.5mL

aliquot and store at -20C, can add 50-100mM DTT instead of BME to completely reduce protein samples