37.5:1 Acrylamide to Bisacrylamide Stabilized Solution
Optimized for SDS-PAGE of Proteins (Laemmli gels)
Consistently Crystal Clear Gels, Zero Fluorescence
Stabilized for Long Shelf Life
Catalog Number: EC-890
ProtoGel forms an electrophoresis matrix that is ideal for the separation of proteins and polypeptides. ProtoGel is a stabilized, ready-to-use 30% (w/v) acrylamide/methylene bisacrylamide solution (37.5:1 ratio), manufactured from the highest quality materials, from which virtually all impurities have been removed. ProtoGel has zero acrylic acid content, eliminating the fixed charges that cause band streaking. Additionally, oxidation products such as aldehydes have been removed by a selective adsorption process. Aldehydes can attack proteins, altering the structure of the sample, and thus altering Rf values. With ProtoGel, you can trust that your results will be consistent one electrophoretic run to the next.
Storage: ProtoGel is stable for 24 months when stored tightly capped in a dark area at room temperature (20°C).
ProtoGel (30%) Protocol
Measure and Mix Solutions:
The volume of ProtoGel required for gel casting solutions of any volume and acrylamide concentration may be calculated from the following formula:
Vp =(X) (Vt)/30
where:
Vp = Volume of 30% ProtoGel
X = % Monomer Desired in Gel
Vt = Total Volume of Gel Casting Solution
EXAMPLE: To make 100 ml of a 10% monomer gel, calculate the volume of Protogel to add as follows:
Vp = (10) (100)/30 = 33.3 ml
Initiate and Cast Gel:
For optimal results degas gel solution for 10 minutes under vacuum aspiration prior to innitiation with APS and TEMED. Add 1.0ml of 10% (w/v) ammonium persulfate for every 100ml of gel casting solution. Swirl gently to mix. Add 0.1 ml of TEMED for every 100ml of gel casting solution. Swirl gently to mix. Pour the solution into the gel casting cassette. The gel should begin to set in 10-20 minutes. To provide a sharp interface, overlay the gel with water saturated n-butanol during polymerization. Flush butanol away with water just before casting the stacking gel (below).
Cast Stacking Gel:
Use ProtoGel Stacking Buffer to make 10ml of a 4% stacking gel:
ProtoGel: 1.3ml
ProtoGel Stacking Buffer: 2.5ml
Deionized Water: 6.1ml
Add 0.05ml 10% Ammonium Persulfate and 0.01ml of TEMED. Gel will begin to set in 20 minutes.