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%①acrylamide/bis mixed solution(37.5 : 1)②
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
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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