10
4.4.4 PreCotes, PreNets
1) Unpack the gels and remove the protective cover-film from the gel surface. Keep
the cover film as it can serve as a protective sheet later.
2) Adjust sample concentration to about 1 - 10 mg protein/ml and desalt by dialysis;
by dilution with dest. water or by lyophilization and resuspension in dest. water.
3) Centrifuge the samples for 5 minutes at approx. 12,000 g; use only the
supernatant. By omitting this step the separation pattern might become fuzzy and,
eventually, precipitates may form within the applicator strip slots.
4) Position the applicator strip on the gel and slightly pressing it with the back of a
forceps. Apply the required sample volume using a pipette. Do not leave empty
slots between samples. Depending on sample type, it is possible to apply the
samples with or without pre-focusing.
4.4.5 CleanGel IEF
1) Unpack and rehydrate the dry gel according to the gel instruction manual.
2) Prepare and apply samples on the gel according to the related gel instruction
manual or specific application procedure.
4.4.6 FocusGel
1) Unpack the gels and remove the protective cover-film from the gel surface. If
necessary, remove excessive moisture from the gel surface with the edge of a
drying cardboard. Keep the cover film as it will serve as a protective sheet later.
The gel is ready to use.
FocusGel 24S and 40S: For some sample types, e.g. serum and CSF the position
of the pre-formed wells is optimized for anodal application in a pH gradient 6-11.
This well position might also be suitable for other sample types. Nonetheless, the
gels can be turned around for cathodal application if needed.
2) Apply the required sample volume using a pipette. Do not leave empty slots
between samples. Depending on sample type, it is possible to apply the samples
with or without pre-focusing.
3) Take care that the electrodes are placed directly on the FocusGel surface at the
gel edges and not on the support film!
DOMINIQUE DUTSCHER SAS
To Next Page
Loading...