1.

Draw a line of origin at one end of the chromatography paper

with a pencil.

2. On the line, add the endoprotease enzyme extract with a fine

needle to give a small spot. Let it dry before adding another drop.
Repeat this until a concentrated spot is obtained.

3. Place the chromatography paper into solvent, making sure the

line of origin is above the solvent surface.

4. Let the chromatography run until the solvent reaches the

solvent front. Cover the container while running the
chromatography to prevent evaporation of the solvent.

5. Let the chromatogram dry before spraying with same specific

dye to locate the products of hydrolysis.

6 Run another 2 replicates for the endoprotease. Make sure the

number of drops of the enzyme extract place on the line of origin
is same as step 2.

7. Repeat step 1 to 5 for the other enzyme, exoprotease. Make

sure the numbers of drops of the enzyme extract place on the line
of the origin is the same as the chromatography for the
endoprotease.

8. For all the chromatograms, measure the distance travelled by

each product in order to calculate the Rf values.

9. The mean Rf values for the chromatograms of both type of the

enzyme is compared.

10. Risk : Solvent easily flammable

Precaution : Make sure there is no naked flame present