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For this next step we will separate the individual proteins in our sample lysate based upon their molecular weight using a positive electrode to attract the negatively charged proteins. To do this we load our previously prepared protein samples into a commercially available polyacrylamide gel. Gels are available in fixed percentages or gradients of acrylamide. The higher the acrylamide percentage the smaller the pore size of gel matrix. Therefore higher percentage gels are better for low weight proteins, low percentage gels are better for large weight proteins and gradient gels can be used for proteins of all sizes due to their varying range in pore size. Prepare your gel by inserting it into the electrophoresis apparatus and filling with running buffer that is appropriate for your gel chemistry. Rinse the wells of the gel with running buffer and add buffer to the chambers. Load your samples into the wells. If you are unsure of the amount to load, 10-30 micrograms of total protein is a
