Hoefer IEF100 - Manual (Page 28)

• p22
Note: Rehydration solutions typically
contain high concentrations of urea
that tend to crystallize at lower
temperatures. For this reason,
rehydration should not be done in cold
rooms or in environments significantly
colder than 20 °C.
Note: Sliding the strips in the
rehydration buffer will wet the entire
length of the strip, and helps prevent
sticking. Strips placed directly against
the plastic bottom of the rehydration
tray may stick, and not rehydrate
properly.
Rehydration of IPG Strips
A brief discussion of rehydration ingredients and their function is
included in Appendix B, along with some recommended solutions.
The reagents can be customized for specific samples.
The volume of solution is dependent on the length of the strip to be
used. Hoefer recommends following the IPG strip manufacturer’s
recommendations.
A table of typical volumes is included below.
Strip Length (cm) Volume per Strip (μl)
7 130
18 340
24 450
Apply rehydration solution in a channel of the appropriately sized
rehydration tray in a line slightly shorter in length than the strip to
be rehydrated.
Many IPG strip manufacturer’s cover the strips with a protective
cover. If present, peel off protective layer.
Place the strip, gel side down, onto the liquid in the channel. Gently
slide the strip back and forth to wet the entire length of the IPG gel
with rehydration solution. Sometimes, lifting and lowering the IPG
strip can aid in getting the proper contact of the solution over the
entire length of the IPG gel strip.
Cover the strips with mineral oil, Hoefer order number GR138-1.
Allow to rehydrate a minimum of 8 hours. Better rehydration occurs
if allowing strips to rehydrate overnight.
Note: Using more than the
recommended volumes may contribute
to loss of protein or blurry IEF results.
Note: The IEF100 is a convenient place
to rehydrate strips. The cold plate will
maintain a constant temperature, and
the cover will help protect the IPG strips
from dust.