Pouring and Running a Protein Gel by reusing Commercial Cassettes

Summary

Our protocol demonstrates how to pour multiple protein gels at a time by recycling Invitrogen Nupage Novex minigel cassettes, and inexpensive materials purchased at a home improvement store. This economical and streamlined method includes a way to store the gels at 4°C for a few weeks. By re-using the plastic gel cassettes from commercially available gels, labs that run frequent protein gels can save significant costs and help the environment.

Abstract

The evaluation of proteins using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is a common technique used by biochemistry and molecular biology researchers^1-4. For laboratories that perform daily analyses of proteins, the cost of commercially available polyacrylamide gels (˜$10/gel) can be considerable over time. To mitigate this cost, some researchers prepare their own polyacrylamide gels. Traditional methods of pouring these gels typically utilize specialized equipment and glass gel plates that can be expensive and preclude pouring many gels and storing them for future use. Furthermore, handling of glass plates during cleaning or gel pouring can result in accidental breakage creating a safety hazard, which may preclude their use in undergraduate laboratory classes. Our protocol demonstrates how to pour multiple protein gels simultaneously by recycling Invitrogen Nupage Novex minigel cassettes, and inexpensive materials purchased at a home improvement store. This economical and streamlined method includes a way to store the gels at 4°C for a few weeks. By re-using the plastic gel cassettes from commercially available gels, labs that run frequent protein gels can save significant costs and help the environment. In addition, plastic gel cassettes are extremely resistant to breakage, which makes them ideal for undergraduate laboratory classrooms.

Protocol

1. Preparing the cassettes

1. Clean front and back plates of a used Invitrogen Nupage Novex minigel cassette, or other commercial minigel cassette.
2. Prepare approximately 1-3 ml of plastic epoxy and mix well according to manufacturer's instructions.
3. Apply the epoxy to the inside edge of the front plate, where the two plates will come in contact when the cassette is assembled.
4. Slide a piece of filter paper or cardboard through the bottom slit of the back plate when assembling the cassette to prevent excess epoxy from sealing the opening. Remove the filter paper as soon as the two plates are assembled.
5. Place binder clips ....

Discussion

Protein gel electrophoresis is an indispensible method for most molecular biology laboratories. Commercially available minigels provide a high level of convenience, but can be costly. Here we show how to re-use the minigel cassettes from a popular brand of commercially available minigels. This method retains the convenience of having a ready-made source of gels, but at a fraction of the cost of commercially available minigels. A key step in this procedure is applying the correct amount of epoxy to completely seal the ca.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Invitrogen Nupage Novex Minigel Cassette (1.0 mm)	Invitrogen	NC2010	Available in 2, 5, 10, 12, 15 and 2D wells
Plastic Epoxy	Loctite	108771
Electrical Tape	Scotch Co.
125 mL side arm flask	Pyrex	5360
4X Tris-Base resolving gel buffer			1.5 M Tris-base pH 8.7
acrylamide	Fisher Scientific	01065-500
Bis-acrylamide	MP Biomedicals	800173
30.8% acrylamide solution			30:0.8 acrylamide: bis-acrylamide in IRG-H2O
10% APS	Fisher Scientific	BP179-25	0.1 g ammonium persulfate in 1 ml IRG-H2O, prepared immediately before use
TEMED	Acros Organics	138450500
1-butanol saturated with 1X resolving gel buffer			5:2 1-butanol to 1X resolving gel buffer
4X Tris-Base stacking gel buffer			1.5 M Tris-base pH 6.8
Gel comb	Invitrogen	NC3015	15 well, 1.0 mm thickness
Impulse Sealer	TEW Electric Heating Equipment
Heat Sealable Pouch	Kapak/Scotchpak		1 pint size (6.5" x 8")
Gel storage buffer			1X stacking gel buffer with 0.1% sodium azide
Novex SDS Tris-Glycine sample buffer (2X)	Invitrogen	LC2676
DTT	Fisher Scientific	BP172-5
10X Tris-Gylcine Running buffer	Invitrogen	LC2675	1L of 10X stock: 29.0 g Tris-Base, 144.0 g Glycine, 10.0 g SDS
XCell SureLock Mini-Cell	Invitrogen	EI0001
Gel Knife	Invitrogen	EI9010
SimplyBlue Safe Stain	Invitrogen	LC6065