How to make SDS Gel?

Tuesday, January 10, 2017

"A very common method for separating proteins by electrophoresis uses a discontinuous polyacrylamide gel as a support medium and sodium dodecyl sulfate (SDS) to denature the proteins. The method is called sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE)."


First, get two gel casts. Each gel cast consists of four gel plates, four gel plastic and eight spacers. Next, Assemble everything and lock them using the gel cast cover. Then, get the acrylamide kit from the refrigerator and place it in fume hood. Excessive inhalation of acrylamide can cause paralysis.
Prepare 10% of resolving gel mix: Where proteins run according to their molecular weight.
Mix the following 38.80mL nano pure water, 19.88mL acrylamide, 20.00mL resolving buffer, 760µl SDS, 560µl APS and 44µl temed were mixed all in beaker. Then carefully shake the solution. Set the pipetor in 5mL and use it to get the prepared solution (resolving gel mix) and placed to the four gel casting. Add 300µl of isoprophanol for the gels to have a smooth edges then set aside these prepared gels and let them solidify for 30 minutes.
Prepare stacking gel mix: Has wells (10-15 wells) that allows the protein to pass through
Mix the following 20.64mL nano pure water, 3.24mL acrylamide, 8.04mL stacking gel buffer, 300µl SDS, 300µl APS, and 40µl temed. These were used and placed at the top of the resolving gel mix and inserted by 10-well comb and 15-well comb. Add stacking gel on top of solidified resolving gel.









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