Pro-PAGE PLUS* - E668  8%, E680  10%, E663  12%

Directions for use

• PAGE-PLUS Concentrate(code E562)

• Pro-PAGE PLUS Buffer(codes J123, J126 or J129)

• Pro-PAGE PLUS 4% Stacking Gel Mix(code E676)

Add the entire contents of one bottle of PAGE-PLUS Concentrate to one bottle of Pro-PAGE PLUS Buffer (the buffer bottle has sufficient volume to allow the addition of one bottle of PAGE-PLUS Concentrate). Cap the bottled mixture tightly and mix gently by inversion - Do Not Shake. This mixture is stable for three months when stored at 4°C. For best results, if not used within three months, discard unused contents by polymerization. Unopened bottles of PAGE-PLUS Concentrate and Pro-PAGE PLUS Buffer are stable for one year when stored at 4°C.

Pro-PAGE PLUS 4% Stacking Gel is premixed and provided for those applications where a stacking gel is desired.

• Determine the total volume of the gel plates you are using. Aliquot the required amount of Pro-PAGE PLUS mixture into a sidearm flask. (Note: It is usually helpful to allow an additional 10 ml to accommodate any possible leaks in the apparatus.) Degas under gentle vacuum for five to ten minutes at room temperature.

• Add Ammonium Persulfate (code 0486) from a 10% stock solution freshly prepared in deionized water at a ratio of 100 l for each 10 ml of Pro-PAGE PLUS mixture, and TEMED (code 0761) at a ratio of 10 L for each 10 ml Pro-PAGE PLUS mixture. If slower polymerization is desired, 60 L of 10% ammonium persulfate solution and 6 L of TEMED may be used, respectively. Mix the solutions by swirling gently, and immediately pour into a prepared glass plate assembly. If a stacking gel will be used, allow resolving gel to polymerize for about 30 min.

• (Optional) Polymerize and pour Pro-PAGE PLUS 4% Stacking gel (code E576) to fill the remainder of the assembly. Insert an appropriately sized comb into assembly and allow gel to polymerize completely (30-45 minutes in most cases). Proceed with electrophoresis in 1X TG-SDS.