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Table of Contents
Page
Acknowledgements……………………………………………………… i
Common Abbreviations List…………………………………………….. ii
List of Figures…………………………………………………………… iii
List of Tables……………………………………………………………. iv
Abstract………………………………………………………………….. 1
Introduction…………………………………………………………… 2 – 10
I. Proteomics …………………………………………………………………. 2
II. The Basis of Separation in Two-dimensional Gel Electrophoresis………… 4
III. Challenges of Two-dimensional Gel Electrophoresis………………………7
VI. Specific Aims……………………………………………………………….8
Methods and Materials………………………………...…………….. 11 – 26
I. Growth conditions…………………………………………………………… 11
II. Centrifugation….…………………………………………………………… 16
III. Sonication………………………………………………………………….. 16
IV. Determining Protein Concentration…………………………………………17
V. Treatment with RNAse………………………………………………………17
VI. Rehydration of IPG strips……………………………………………………17
VII. Isoelectric Focusing (IEF)…………………………………………………. 18
VIII. Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis…………… 19
IX. Staining the gel………...……………………………………………………21
X. Image Analysis………………………………………………………………22
XI. Analysis with Phoretix 2D Software………………………………………...22
Results and Discussion…………………………………………….… 27 – 52