Biology Reference
In-Depth Information
20. Wells are separated between samples to prevent mixing up
samples and electrophoresis. Electric power supply is turned
on constant voltage mode.
21. Be careful not to mix with the next sample bands. Illuminate
the glass plate from below with a fl uorescent lamp to see the
gels easily.
22. At this stage, dehydrated, compressed, and completely
bleached gels should be observed. If the gels do not change,
repeat this step twice.
23. At this stage, rehydrated and swollen gels should be observed.
If the gels do not change, repeat this step twice.
24. Gels are sometimes partly bleached, but this is acceptable.
25. At this stage, dehydrated and compressed gels are easily lost by
electrostatic force. Take extra care.
26. Digested and purifi ed peptides should be analyzed by nano-
LC-MS/MS within 1 week.
Acknowledgements
This work was supported in part by Grants-in-Aid for Scientifi c
Research (#22120003 and #24370018) from MEXT, Japan, to
Y.K. and M.U.
References
1. Aebersold R, Mann M (2003) Mass spectrom-
etry-based proteomics. Nature 422:198-207
2. Domon B, Aebersold R (2006) Mass spec-
trometry and protein analysis. Science
312:212-217
3. Kersten B, Burkle L, Kuhn E-J et al (2002)
Large-scale plant proteomics. Plant Mol Biol
48:133-141
4. van Wijk KJ (2001) Challenges and prospects
of
8. Rabilloud T (2009) Membrane proteins and
proteomics: love is possible, but so diffi cult.
Electrophoresis 30:174-180
9. Rabilloud T (2002) Two-dimensional gel elec-
trophoresis in proteomics: old, old fashioned,
but it still climbs up the mountains. Proteomics
2:3-10
10. Simons K, Ikenen E (1997) Functional rafts in
cell membranes. Nature 387:569-572
11. Peskan T, Westermann M, Oelmuller R (2000)
Identifi cation of low-density Triton X-100-
insoluble plasma membrane microdomains in
higher plants. Eur J Biochem 267:6989-6995
12. Mongrand S, Morel J, Laroche J et al (2004)
Lipid rafts in higher plant cells. J Biol Chem
279:36277-36286
13. Bhat RA, Panstruga R (2005) Lipid rafts in
plants. Planta 223:5-19
14. Martin SW, Glover BJ, Davies JM (2005)
Lipid microdomains: plant membranes get
organized. Trends Plant Sci 10:263-265
15. Grennan AK (2007) Lipid rafts in plants. Plant
Physiol 143:1083-1085
16. Thakur S-S, Geiger T, Chatterjee B et al
(2011) Deep and highly sensitive proteome
plant
proteomics.
Plant
Physiol
126:501-508
5. Santoni V, Kieffer S, Desclaux D et al (2000)
Membrane proteomics: use of additive main
effects with multiplicative interaction model to
classify plasma membrane proteins according
to their solubility and electrophoretic proper-
ties. Electrophoresis 21:3329-3344
6. Luche S, Santoni V, Rabilloud T (2003)
Evaluation of nonionic and zwitterionic deter-
gents as membrane protein solubilizers in two-
dimensional
electrophoresis.
Proteomics
3:249-253
7. Gorg A, Weiss W, Dunn M-J (2004) Current
two-dimensional electrophoresis technology
for proteomics. Proteomics 4:3665-3685
