Identification of Tumor-Associated Antigens 9
The IEF strips were overlayered with a solution containing
agarose (0.5% w/v) on the top of second-dimension gels. The
second dimension was carried out using 10% or 15% polyacryla-
mide gels and current is nonlimited per gel in a cooling system.
8. Spot patterns on each membrane are matched to an equivalent
spot on original 2D-PAGE gel. This gel analysis can be done
by using PDQuest 2-D Analysis Software (Bio-Rad, Hercules,
CA).
9. If the gel pieces still have Coomassie blue staining, the gel
pieces need to be rehydrated again in an equal volume of
ammonium bicarbonate and acetonitrile.
This work was supported by grants from National Institutes of
Health (2S06GM008012-37, 5G12RR08124, CA56956).
Acknowledgments
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