Hoefer SE900 - Manual (Page 20)

• p14
Disassembling the Multiple Gel Caster
1 Place the caster down on its back in a tray or sink.
2 Remove the red clamps and screws securing the faceplate to
the caster.
3 Slide off the faceplate.
4 Remove gel cassettes and filler sheets. Rinse outside of the gel
cassettes to remove excess polymerized acrylamide.
5 Clean the multiple gel caster components with laboratory detergent.
Rinse and let air dry.
IPG Strip Equilibration
Before IPG strips are placed on top of the second dimension gel, the
buffer in the strips needs to be replaced with an appropriate buffer
for PAGE.
Equilibrate IPG strips in appropriate equilibration buffers. Typically,
a two step equilibration (first with DTT and a second with iodoacet-
amide) gives better results than a single DTT equilibration step. The
following procedure is recommended.
Equilibration Procedure
1 Thaw two aliquots of the equilibration solution.
2 Add 10 mg/ml DTT to one solution.
3 Place the IPG strips in the rehydration/equilibration tray, or a
glass tube.
4 Add 6.5 ml of solution to each slot containing an IPG strip.
5 Place on rocker for 10–15 minutes.
Following equilibration, discard the first equilibration solution in an
appropriate manner.
6 Add 25 mg/ml iodoacetamide (IAA) to the second aliquot of
equilibration solution.
7 Add 6.5 ml of solution to each slot containing an IPG strip.
8 Place on rocker for 10–15 minutes.
Following equilibration, discard the second equilibration solution in an
appropriate manner.
Following equilibration, the IPG strips are placed on the top of the second
dimension gel, and sealed into place with the agarose overlay.
Note: The Plate Rack, SE914, is a
convenient accessory for temporarily holding
the gel cassettes when disassembling the
multiple gel caster.