1. Pensacola High School IB Chemistry 3
Gel Electrophoresis
Pensacola High School
IB Chemistry 3

2. Steps Breakdown the protein into amino acids
Break amide/peptide bond
Called hydrolysis (opposite of condensation)
Acid or base and heat required
Add protein to gel containing a buffer
Apply a voltage
Amino acids move based on mass and charge
Dye amino acids
Ninhydrin
Measure distance traveled,
Compare to known

3. Set-up

4. Why amino acids move +NH3-C-COO-
Amino acids separate based on their isoelectric point and molar mass
Isoelectric point:
This is the pH where they net charge of amine and carboxylic acid groups cancel out
+NH3-C-COO-

5. pH of buffer is more basic than isoelectric point, than amino acid will have a negative charge and move toward postive electrode
There are fewer hydrogen atoms around to protonate it
NH2-C-COO-
pH of buffer is more acidic than isoelectric point, than amino acid will have a positive charge and move to negative electrode
More H atoms around to protonate
+NH3-C-COOH