ProteomIQ blue, a potent post-stain for the visualization and subsequent mass spectrometry based identification of fluorescent stained proteins on 2D-gels.
Manual spot excision for protein identification from fluorescent stained two-dimensional (2-D) gels is hard to accomplish. Here, we explore the use of ProteomIQ Blue as a post-stain method for the visualization of fluorescent stained/labeled proteins. We show that ProteomIQ Blue post-staining is almost as sensitive as staining with SYPRO Ruby or cyanine dyes alone. More than 90% of the protein spots that are stained with the fluorescent stains are still detectable with ProteomIQ Blue. In protein identification by mass spectrometry, ProteomIQ Blue post-stained spots provide high sensitivity and high protein sequence coverage of the peptide mass maps in both MALDI-TOF-MS and ESI-MS/MS analyses. In conclusion, post-staining of fluorescent stained gels with ProteomIQ Blue provides a facile and a powerful method to achieve quantitative protein analysis as well as protein identification in the same semianalytical gel without requiring sophisticated/expensive robotic equipment.
Wijte
Dorien
D
TNO Defence, Security and Safety, P.O. Box 45, 2280 AA Rijswijk, The Netherlands. dorien.wijte@tno.nl
de Jong
Ad L
AL
Mol
Marijke A E
MA
van Baar
Ben L M
BL
Heck
Albert J R
AJ
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002232:Carbocyanines / Q000737:chemistry; D004396:Coloring Agents / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D009942:Organometallic Compounds / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013194:Staining and Labeling / Q000379:methods*
D001426:Bacterial Proteins; D002232:Carbocyanines; D004396:Coloring Agents; D005456:Fluorescent Dyes; D009942:Organometallic Compounds; C515601:Sypro Ruby