SDS-PAGE Gel Calculator

How to make SDS-PAGE gels

1. Enter the resolving gel percentage and how much gel each plate holds, and the same for the stacking gel.
2. Enter the number of gels you are pouring.
3. The table lists the water, acrylamide, Tris, SDS, APS and TEMED for each gel layer. Add APS and TEMED last, just before pouring.

Frequently asked questions

Which recipe does it use?

Laemmli gels from a 30% acrylamide/bis stock, with 0.375 M Tris pH 8.8 in the resolving gel and 0.125 M Tris pH 6.8 in the stacking gel, 0.1% SDS and 0.1% APS. TEMED is 0.04% in the resolving gel and 0.1% in the stacking gel.

Which percentage should I use?

Higher percentages separate smaller proteins. Roughly: 8% for about 40 to 200 kDa, 10% for 20 to 150 kDa, 12% for 15 to 100 kDa and 15% for 10 to 70 kDa.

How much gel does a plate take?

It depends on the casting system and spacer thickness. A typical 1.0 mm mini gel takes about 5 mL of resolving gel and 2 mL of stacking gel; 1.5 mm plates take about half as much again.

Is acrylamide dangerous?

Unpolymerized acrylamide is a neurotoxin. Wear gloves, and handle the stock and unset gel solutions with care.