Amylase in animals, plants and bacteria can be detected and characterized by the procedure described in this exercise. The analysis is performed by incorporating starch into an SDS-agarose gel prior to electrophoresis of the denatured proteins. After electrophoresis, the SDS is removed and the enzyme spontaneously renatures. The location of amylase in the gel is then carried out by staining the starch in the gel with iodine. Zones of enzyme activity are devoid of starch and are seen as clear bands against a background of blue. Using this procedure, students determine the molecular weight of amylase in their own saliva and compare this value to the molecular weight of amylases produced by bacteria.
Each of the individual experiments are supplied with the chemicals and laboratory guides needed for 16 students working in pairs. If you chose one or more of the experiments below – you should also order Electrophoresis Package 2M or Electrophoresis Package 2. Electrophoresis Package 2M provides sufficient agarose, gel stain, and electrophoresis buffers for 1 of the individual experiments in this series (four gels with 15ml of agarose per gel). Electrophoresis Package 2 provides sufficient agarose, gel stain, and electrophoresis buffers for up to 6 of the individual experiments in this series.