2D DIGE Analysis of Protein Extracts from Muscle Tissue - Difference Gel Electrophoresis (DIGE): Methods and Protocols - page 167

Table 1
Excitation and emission characteristics for the three
common DIGE fl uors

Laser excitation
source (nm) Fluorescent dye Emission fi lter (nm)

DIGE fl uor Cy3 minimal dye Green (532) 580 BP 30

DIGE fl uor Cy5 minimal dye Red (633) 670 BP 30

DIGE fl uor Cy2 minimal dye Blue (488) 520 BP 40

Acknowledgements

This work was supported by the Telethon foundation (grant
N. GGP08107D to C.G.), EU community (grant BIO-NMD N.
241665 to C.G.), and Italian Ministry of University and Scientifi c
Research (grant FIRB RBRN07BMCT to C.G.).

References

1. Unlu, M., Morgan, M. E., and Minden, J. S.
(1997) Difference gel electrophoresis: a single
gel method for detecting changes in protein
extracts, Electrophoresis 18 , 2071-2077.
2. Garrels, J. I. (1979) Two dimensional gel elec-
trophoresis and computer analysis of proteins
synthesized by clonal cell lines, J Biol Chem
254 , 7961-7977.
3. Klose, J. (1975) Protein mapping by combined
isoelectric focusing and electrophoresis of
mouse tissues. A novel approach to testing for
induced point mutations in mammals,
Humangenetik 26 , 231-243.
4. O'Farrell, P. H. (1975) High resolution two-
dimensional electrophoresis of proteins, J Biol
Chem 250 , 4007-4021.
5. Gharbi, S., Gaffney, P., Yang, A., Zvelebil, M. J.,
Cramer, R., Waterfi eld, M. D., and Timms, J. F.
(2002) Evaluation of two-dimensional differ-
ential gel electrophoresis for proteomic expres-
sion analysis of a model breast cancer cell
system, Mol Cell Proteomics 1 , 91-98.
6. Zhou, G., Li, H., DeCamp, D., Chen, S., Shu, H.,
Gong, Y., Flaig, M., Gillespie, J. W., Hu, N.,
Taylor, P. R., Emmert-Buck, M. R., Liotta, L.
A., Petricoin, E. F., 3 rd, and Zhao, Y. (2002)
2D differential in-gel electrophoresis for
the identifi cation of esophageal scans cell

cancer-specifi c protein markers, Mol Cell
Proteomics 1 , 117-124.
7. Swatton, J. E., Prabakaran, S., Karp, N. A.,
Lilley, K. S., and Bahn, S. (2004) Protein pro-
fi ling of human postmortem brain using
2-dimensional fl uorescence difference gel
electrophoresis (2-D DIGE), Mol Psychiatry 9 ,
128-143.
8. Alban, A., David, S. O., Bjorkesten, L.,
Andersson, C., Sloge, E., Lewis, S., and Currie,
I. (2003) A novel experimental design for com-
parative two-dimensional gel analysis: two-
dimensional difference gel electrophoresis
incorporating a pooled internal standard,
Proteomics 3 , 36-44.
9. Knowles, M. R., Cervino, S., Skynner, H. A.,
Hunt, S. P., de Felipe, C., Salim, K., Meneses-
Lorente, G., McAllister, G., and Guest, P. C.
(2003) Multiplex proteomic analysis by two-
dimensional differential in-gel electrophoresis,
Proteomics 3 , 1162-1171.
10. Marouga, R., David, S., and Hawkins, E.
(2005) The development of the DIGE system:
2D fl uorescence difference gel analysis technol-
ogy, Anal Bioanal Chem 382 , 669-678.
11. Yan, J. X., Devenish, A. T., Wait, R., Stone, T.,
Lewis, S., and Fowler, S. (2002) Fluorescence
two-dimensional difference gel electrophoresis