2-D Electrophoresis Using Immobiline Drystrip - Pharmacia Biotech Multiphor II User Manual

5. Operation
5.7 2-D electro-
phoresis
using
Immobiline
DryStrip and
ExcelGel SDS
gradient
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This chapter gives a brief description of the 2-D (two dimensional)
electrophoresis method using Immobiline DryStrip and ExcelGel SDS. For
more detailed information on running conditions, please refer to the
instructions supplied with Immobiline DryStrip Kit.
Isoelectric focusing using Immobiline DryStrip makes true isoelectric
focusing possible and significantly improves the reproducibility of the spot
distribution along the pH gradient axis of 2-D maps. Immobiline DryStrip
also makes it possible to obtain distinct protein spots, even of basic
proteins.
The Immobiline DryStrip Kit facilitates sample application, running and
equilibration of Immobiline DryStrip for the first dimension of 2-D
electrophoresis. The kit includes the accessories necessary to run up to 12
Immobiline DryStrip strips simultaneously on Multiphor II. The high
sample capacity allows the application of up to 100 µl on each Immobiline
DryStrip.
First dimension:
Immobiline DryStrip are precast polyacrylamide gels (T=4%, C=3%)
containing an immobilized pH gradient, cast on plastic support film.
The strips are delivered in a dehydrated form. After rehydration, the gel
thickness of the strips is 0.5 mm.
Immobiline DryStrip gels are available in different sizes (110 x 3 mm and
180 x 3 mm) and pH ranges (4-7 and 3-10). The shapes of the pH
gradients are described in the following table.
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