Reagents for preparing polyacrylamide gel for SDS-PAGE

Examples of polyacrylamide gel preparation for SDS-PAGE are summarized by each gel concentration. Please use an acrylamide/bis mixed solution with an acrylamide/bis ratio of 37.5:1 or 29:1.

Amounts of reagents required to prepare separation gel

For 10 mL of separation gel
Gel concentration (%T)		6%		8%		10%		12%		15%
Concentration of acrylamide/bis mixed solution		30w/v%	40w/v%	30w/v%	40w/v%	30w/v%	40w/v%	30w/v%	40w/v%	30w/v%	40w/v%
Product Number	Product Name
043-16785	Water(mL)	5.4	5.9	4.7	5.4	4.1	4.9	3.4	4.4	2.4	3.65
-	Acrylamide/bis mixed solution (mL)	2.0	1.5	2.7	2.0	3.3	2.5	4.0	3.0	5.0	3.75
192-11041	Separating Gel Buffer Solution (x 4)	2.5	2.5	2.5	2.5	2.5	2.5	2.5	2.5	2.5	2.50
019-15922	10w/v% APS(mL)	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.10
	Total volume (mL)	10.0	10.0	10.0	10.0	10.0	10.0	10.0	10.0	10.0	10.00
208-17452	TEMED(mL)	0.008	0.008	0.006	0.006	0.004	0.004	0.004	0.004	0.004	0.004

* Deaeration is recommended before addition of TEMED.
* When TEMED is added, the gel starts to polymerize. Perform subsequent operations quickly.

Amounts of reagents required to prepare concentrated gel

5% concentrated gel
Concentration of acrylamide/bis mixed solution		30w/v%	40w/v%	30w/v%	40w/v%
Product Number	Product Name	For 1 mL		For 5 mL
043-16785	Water(mL)	0.690	0.735	3.490	3.695
-	Acrylamide/bis mixed solution(mL)	0.170	0.125	0.830	0.625
199-11051	Concentrated Gel Buffer Solution (x 4)	0.130	0.130	0.630	0.630
019-15922	10w/v% APS(mL)	0.010	0.010	0.050	0.050
	Total(mL)	1.000	1.000	5.000	5.000
208-17452	TEMED(mL)	0.001	0.001	0.005	0.005

Modified from Joseph and David. Cold Spring Harb Protoc; 2006.