Question

In the cell bio lab, we use company manufactured gels, however you can make you own polyacrylamide gels. List all of the ingredients found in an SDS-PAGE gel. Which ingredients are responsible for polymerizing the solution?

Answer 1

The list of ingredients are used in this method are as follow-

1. Acrylamide - mostly 30%

2. Bisacrylamide - mostly 0.8%

3. 1.5M Tris-Hcl pH 6.8 for stacking gel and 8.8 for separating gel.

4. 10% SDS

5. 10% APS

6. TEMED

7. Double distilled water

Polymerization:

Acrylamide is a polymer that is used to dissolved in buffer and bisacrylamide is the cross linking agen.

Two types of gels are prepared for separation of protein. Stacking gel is usually 4 or 6% and separating gel is usually 10 to 20%. Lower concentration of stacking gel will allow the migration of all the protein same because of larger pore size. It will stack or line up the proteins for its next level of separation.

Higher concentration of separating gel will provide resistance to the movement of protein because of the low pore size and therefore linear bands can be achieved neatly.