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10. Spot detection and matching can also be performed using the
“batch processor” of the software simultaneously for all gels.
11. To avoid a high sample volume for the preparative labeling,
TCEP and Cy3 should have a concentration of 20 mM instead
of 2 mM. The amount of TCEP and Cy3 for labeling of the
preparative protein lysate has to be calculated according to
labeling conditions for analytical gels.
12. Before scanning, mark the glass plate from the picking gel with
fl uorescence stickers. The stickers are necessary for matching
the gel as the proteins are not visible and the gel image. After
scanning, print the gel image in the original size. Put the print
under the glass plates and align the position of the stickers on
the glass plate with the image.
Acknowledgments
The authors would like to thank Anna Lendzian and Sabine
Roggenbrodt for excellent technical assistance. This work was sup-
ported by a grant from the European Commission (LCVAC,
COOP-CT-2004-512855).
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