Get exact volumes for Tris-Glycine (Laemmli) resolving and stacking gels at any acrylamide percentage.
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Select your desired resolving gel percentage based on the size of your target protein(s), choose the number of gels and gel format, and the calculator provides exact volumes for both the resolving and stacking gels. APS and TEMED are added last — they initiate polymerization, so work quickly after adding them.
This calculator uses the standard Tris-Glycine (Laemmli) discontinuous buffer system with 30% acrylamide/bis-acrylamide (29:1 ratio), 1.5 M Tris-HCl pH 8.8 for the resolving gel, and 0.5 M Tris-HCl pH 6.8 for the stacking gel. SDS concentration is 10% (w/v) stock.
Preparation tips: Degas the acrylamide solution briefly before adding APS and TEMED. Pour the resolving gel first, overlay with water-saturated butanol or isopropanol, wait 30–45 minutes for polymerization, rinse off the overlay, then pour the stacking gel and insert the comb. The stacking gel polymerizes in ~15 minutes.
Safety: Unpolymerized acrylamide is a neurotoxin. Always wear gloves when handling acrylamide solutions and dispose of waste properly. Never pour unpolymerized acrylamide down the drain — add excess APS/TEMED to polymerize waste before disposal.