ROLE OF ATG POLYCLONAL ANTIBODIES(10)

The protein samples are the mixed with treatment buffer in the ratio 2:1 and boiled for 5 minutes. Appropriate amount of sample is then loaded into the wells using a microliter syringe. The entire unit is connected to a power pack and a current of 12mA is made to pass through the gel by applying potential difference between the two electrodes. The electrophoresis is preferably run at 40C. This is to counter the heat is produced during the electrophoresis, which might cause the gel to melt. When the proteins enter the separating gel the current is raised to 13 mA. When the electrophoresis completed the gel is stained.

Procedure for Staining in SDS PAGE

Place the gel ready to be stained in distilled water (D/W) for 2 hours. This will be lead to diffusion as unreacted. Proteins and salts into the D/W and will also help in separating the gel from the glass plate. T ake a gelatin paper and cut it to size that is little excess to one size of the plate. Wet the gelatin paper and cover the glass plate. Avoid the trapping of air bubbles. After 2 hours the gel will slip from the slide and will float in water. Remove the glass plate and slip carefully the glass plate covered with gelatin paper under the gel.