ProteomeLab™ PF 2D Analysis of Genetically-Characterized Human Glioma Xenografts
Victoria Ioffe1, Ashken Movsisyan1, Jiuhong Yu1, C. David James2, and Oliver Bögler1
- William and Karen Davidson Laboratory, Hermelin Brain Center, Department of Neurosurgery, Henry Ford Hospital, Detroit, MI
- Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN
Results and Discussion
Proteome maps were generated by the ProteomeLab PF 2D system which separates proteins according to pI in a first-dimension chromatofocusing step and according to hydrophobicity in a reversed-phase step in the second dimension. The data was displayed as a UV/pI map using ProteoVue† software (Figure 1A and B), which displays the pI fractions as lanes and the second dimension retention time as vertical position in individual lanes. The intensity of the bands in the map corresponds to absorbance at 214nm. Lanes 5 to 20 corresponds to the pH range 4.0 to 8.5, which represents the pH gradient in the first dimension. The pH range of each fraction is shown along the top of the windows in ProteoVue. In addition the software has been used to zoom in to show retention times between 6 and 26 minutes in the second dimension where most peaks are found. The peak trace shown at the left side of each panel is from the third lane from the left in each case. This fraction has a pH range of 4.5-4.8. In this fraction, both samples show a major peak around 17 minutes but are otherwise quite different.
| A |  |
| B |
 |
| Figure 1. UV/pI maps of glioblastoma xenograft proteomes in ProteoVue. Total proteomes of two glioblastoma xenografts fractionated with the ProteomeLab PF 2D system are displayed in ProteoVue software. Shown are (A) glioblastoma #6, which carries a ∆EGFR amplification, and (B) glioblastoma #12, which carries an EGFR amplification. | |
† ProteoVue and DeltaVue are trademarks of Eprogen, Inc.
* All trademarks are the property of their respective owners. Where applicable, the PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffman-LaRoche, Ltd.
