Vertical electrophoresis device
Electrophoresis is one of the most well-known laboratory techniques for separating biomolecules, and refers to the movement of a charged body in an electric field at a given pH. In this method, the movement of particles (samples with electric charge) in a liquid under the influence of a uniform electric field through a porous or mesh network is examined.
Vertical electrophoresis is a method for analyzing proteins by electrophoretic separation (polyacrylamide gel). This method is also known as sodium dodecyl sulfate polyacrylamide gel electrophoresis. Polyacrylamide gels restrict the movement of large molecules.
To make a vertical electrophoresis device, the following items are needed:
1. Tank chamber
2. Two glass plates (to make the gel)
3. Spacers
4. Gel-separating comb
5. Solutions needed to make the gel
6. 7 large, stiff clamps
First, we place two glass plates parallel to each other. We separate them using spacers. Then we tighten them on the two sides and bottom using 3 clamps on each side.
To prevent the acrylamide gel we will make from leaking through the pores of the glass plates, it is best to first seal any gaps using an agarose gel.
Refer to the relevant post for instructions on making the agarose gel.
While the agarose gel is still warm and not set, it is best to introduce it into the chamber from the corner sides so that it coats just the walls and does not fill the middle space.
After this, we let some time pass for the gel to harden and set.
Meanwhile, we prepare the acrylamide gel:
To make the acrylamide gel, we need:
1. Stock 30 - 4cc
2. TBE 5x - 4cc
3. H2O - 12cc
4. APS 10% - 300 μl
5. TEMED 25% - 25 μl
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