Dear Dr. Hess;
I am the section head of proteomics at Molecular Probes Incorporated and
responsible for the R&D efforts for the SYPRO dye product line. First, SYPRO
Orange dye is NOT as sensitive as the better silver stains, it has sensitivity
comparable to colloidal Coomassie Blue dye and some of the rapid, lower
sensitivity silver stain kits. SYPRO Red dye and the nonfixative stain,
SYPRO Tangerine dye, also deliver sensitivity that is comparable to colloidal
Coomassie Blue dye. The three dyes are all fully compatible with MALDI-TOF
mass spectrometry, however and provide better sequence coverage and yields
than silver stain. Staining is rapid, complete in only 30-45 minutes. The
SYPRO Ruby dye stains offer detection sensitivity that matches the best silver
stains and are also fully compatible with MALDI-TOF mass spectrometry, Edman
sequencing, amino acid compositional analysis, and other common downstream
techniques. If sensitivity is most important to you, I recommend these
products over SYPRO Orange dye. Staining requires 3 hours to over night for
the gel stains, so if speed counts, SYPRO Orange dye is the best choice.
Recent publications on many of the SYPRO dyes that detail compatibility with
imaging devices and modern microchemical characterization procedures are listed
below:
Berggren, K., Chernokalskaya, E., Steinberg, T., Kemper, C., Lopez, M., Diwu,
Z., Haugland, R. and Patton, W. (2000) Background-Free, High-Sensitivity
Staining of Proteins in One- and Two-Dimensional Sodium Dodecyl
Sulfate-Polyacrylamide Gels using a Luminescent Ruthenium Complex.
Electrophoresis (In Press)
Steinberg, T., Lauber, W., Berggren, K. Kemper, C., Yue, S. and Patton, W.
(2000) ģFluorescence Detection of Proteins in SDS-Polyacrylamide Gels using
Environmentally Benign, Non-Fixative, Saline Solutionī. Electrophoresis 21,
497-508
Berggren, K., Steinberg, T., Lauber, W., Carroll, J., Lopez, M.,
Chernokalskaya, E., Zieske, L., Diwu, Z., Haugland, R. and Patton, W. (1999)
ģA luminescent ruthenium complex for ultrasensitive detection of proteins
immobilized on membrane supportsī. Anal. Biochem. 276: 129-143.
Steinberg, T., Chernokalskaya, E., Berggren, K., Lopez, M., Diwu, Z., Haugland,
R. and Patton, W. (2000) ģUltrasensitive fluorescence protein detection in
isoelectric focusing gels using a ruthenium metal chelate stainī.
Electrophoresis 21, 486-496
Best Regards,
Wayne F. Patton, Ph.D.
Daniel Hess wrote:
> I just saw an advertisement for SYPRO and I am wondering if anybody did try
> SYPRO Orange for protein analysis (Maldi, NanoESI-MSMS)? Is it really
> better in sensitivity than silver staining? How is the peptide recovery
> after tryptic cleavages? Basically is it worthwhile to try?
> Thanks for the info!
> With best regards Daniel
>
> Dr. Daniel Hess
> Friedrich Miescher Institut
> Maulbeerstr. 66
> CH-4058 Basel
> Switzerland
>
> Phone: Office ++41 61 697 85 44, Lab ++41 61 697 68 68
> FAX: ++41 61 697 39 76 e-Mail: dhess@fmi.ch
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