How do I run SDS gel?
Keeping this in view, how do SDS PAGE gels work?
SDS-PAGE separates proteins primarily by mass because the ionic detergent SDS denatures and binds to proteins to make them uniformly negatively charged. Thus, when a current is applied, all SDS-bound proteins in a sample will migrate through the gel toward the positively charged electrode.
Beside this, how do I make acrylamide gel?
Mix acrylamide/bis solution, buffer and water in separate beakers. Deaerate the solutions briefly (1 to 3 min ad vacuo). Add to separating gel solution: 10 % SDS solution (w/v in water), TEMED and APS solution (w/v 10 % of ammonium persulfate), gently swirl to mix without incorporating air into the mixture.
Typical conditions include runs at 100-150 volts for 40-60 minutes or until the dye front has reached the bottom of the gel. Letting it run too long will result in losing your lower molecular weight bands.