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Protein Gel Box
SOP for Owl Scientific Protein Gel Electrophoresis System
Casting the Gel
- Arrange two side spacers between two glass plates, one blank, one notched.
- Place into a gel pouch. Use one bag for each gel.
- Stand glass and bag assembly up in casting stand so that it rests against the rear wall.
- Insert spacer plate in front of the glass and bag assembly.
- Tighten two thumbscrews on the front of the unit just enough to hold the assembly against the rear wall of the casting stand.
- Slide the Spacer Placer card between glass plates to aid in aligning spacers. Remove the Spacer Placer.
- Use the two thumbscrews at the front of the unit to level the casting stand. Observe the bubble level.
- Use the two rear thumbscrews to stabilize the stand so that it does not rock.
- Pour gels from top between glass plates.
- Insert comb, teeth-side down, into top of gel. Allow gel to polymerize.
- Remove gel from bag for immediate use or heat seal top of bag for storage.
Running the Gel
- Use polyacrylamide gels cast from the Joey Gel Casting System above.
- Place upper buffer chamber (UBC) into lower buffer chamber (LBC) so that the pins lock.
- Loosen knobs and place gel assembly on the two orange "feet" at base of the UBC. Offset or eared glass should face center of UBC. (Alumina plate may be placed between glass plate and center of UBC for additional cooling.)
- Slide clamps over plates and finger-tighten knobs to form seal between gasket and plate.
- Repeat for second get, or clamp two blank glass plates onto other side if running only one gel.
- Add buffer to LBC to about 2-3mm above base of gel.
- Add buffer to UBC to about 5mm from top of gel assemblies. Make sure level is above gap formed by "ears". Put lid on.
- Attach 3/8" ID tubing to each nozzle on UBC marked IN and OUT. Attach tubing on IN (black electrode) side to cold water tap. Put ends of tubing on OUT (red electrode) side into sink to drain.
- Prime the system by turning on water enough to circulate through system. Turn off.
- Load samples onto gel. Turn on water. Remember to put safety lid back on.
- When water has started to circulate through system, connect power cords to power supply and begin the run.
- When finished, slide and lift UBC from LBC and drain buffer chambers.
- Remove gel and proceed with analysis.