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Reagents for preparing polyacrylamide gel for SDS-PAGE

Quick reference -Examples of gel preparation 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.

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Acrylamide/bis Mixed Solution

With SDS-PAGE, a solution with an acrylamide/bis ratio of 37.5:1 or 29:1 is usually used. As the bis ratio increases, gel strength and separation ability in the low molecular weight region increase, but the separation ability in the high molecular weight region decreases.

Numbers included in product names

30 w/v%acrylamide/bis mixed solution(37.5 : 1)

① Total concentration of acrylamide and bis
② Mixture ratio of acrylamide: bis

Acrylamide

Cross-linkers (bis)

Gel Buffers

SDS

Tris-HCl

Tris

Hydrochloric Acids

Running Buffers

Sample Buffer (without Reducing Agent)

Sample Buffers (with Reducing Agent)

Reducing Agents

Product content may differ from the actual image due to minor specification changes etc.

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