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5. Operation
Immobiline applicator strip for up to 5 µl of sample in 52 slots. Immobiline
applicator strip is designed to counteract lateral band spreading.
Sample application pieces hold approximately 20 µl of sample. For smaller
volumes, cut the paper pieces to an appropriate size. At least 24 application
pieces size 5 x 10 mm and 50 application pieces size 2.5 x 5 mm can be
placed on one gel. Apply the sample about 1 cm away from the cathodic
buffer strip and 1 cm away from each short side of the gel. For the best
results, remove the application pieces 15 min after electrophoresis has
started.
ExcelGel SDS, ExcelGel Homogeneous 7.5, 12.5, 15 and CleanGel are
available with various numbers of sample wells for various volumes.
Samples are applied directly into the wells immediately prior to the run.
Electrophoresis
Connect Multiphor II to MultiTemp II thermostatic circulator. Switch on
MultiTemp II 15 minutes before starting the experiment and set the
temperature to 15 °C.
Always wear clean gloves when working with polyacrylamide gels and
buffer strips, particularly when using sensitive staining methods.
Remove one ExcelGel SDS from the package. Pipette 1 ml of insulating fluid
(kerosene or light paraffin oil) onto the cooling plate. Place the gel with the
stiff plastic film facing down in the middle of the cooling plate, making sure
no air bubbles are trapped under the gel. Position the gel so that the polarity
of the gel corresponds to that of the plate. Use the screened template on the
cooling plate to centre the gel. Remove any excess solution with paper
tissue. Remove the protective cover film from the gel.
Open the ExcelGel SDS strip packages and apply the cathodic and the
anodic SDS buffer strips on the respective sides of the gel.
Note: The narrowest side of the buffer strip should be placed on the gel
surface.
Choose an appropriate sample application method and apply the sample.
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