Vertical Gel Instrument- The schematic diagram of a vertical gel electrophoresis apparatus is given in Figure 13.3 . It has two buffer chamber, upper chamber and a lower chamber. Both chamber are fitted with the platinum electrodes connected to the external power supply from a power pack which supplies a direct current or DC voltage. The upper and lower tank filled with the running buffer is connected by the electrophoresis gel casted in between two glass plates (rectangular and notched). There are additional accessories needed for casting the polyacrylamide gel such as comb (to prepare different well), spacer, gel caster etc.

Figure 13.3 : Different components of vertical gel electrophoresis apparatus.

Buffer and reagent for electrophoresis- The different buffer and reagents with their purpose for vertical gel electrophoresis is as follows-

1. N, N, N', N'-tetramethylethylenediamine (TEMED)- it catalyzes the acrylamide polymerization.

2. Ammonium persulfate (APS)- it is an initiator for the acrylamide polymerization.

3. Tris-HCl- it is the component of running and gel casting buffer.

4. Glycine- it is the component of running buffer.

5. Bromophenol blue- it is the tracking dye to monitor the progress of gel electrophoresis.

6. Coomassie brilliant blue R250- it is used to stain the polyacrylamide gel.

7. Sodium dodecyl sulphate- it is used to denature and provde negative charge to the protein.

8. Acrylamide- monomeric unit used to prepare the gel.

9. Bis-acrylamide- cross linker for polymerization of acrylamide monomer to form gel.