Author Topic: Flamingo, Sypro Ruby and Deep Purple  (Read 29783 times)

luigi.villa

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Flamingo, Sypro Ruby and Deep Purple
« on: December 13, 2006, 05:00:00 PM »
1)Which is the best in terms of cost/quality ratio?
2)Which is the best for MALDI-TOF analysis?
3)Are they all suitable for post-staining of CyDye stained gels?
Please, answer according to pratical lab experience. Thanks  

Reiner Westermeier

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Re: Flamingo, Sypro Ruby and Deep Purple (luigi.villa)
« Reply #1 on: December 13, 2006, 05:00:00 PM »
Check out these papers:
1) Laane B, Panfilov O,  Protein staining influences the quality of mass spectra obtained by Peptide mass fingerprinting after separation on 2-D gels. A comparison of staining with coomassie brilliant blue and sypro ruby.  J Proteome Res 4 (2005) 175 iV 179.
2) Coghlan DR, Mackintosh JA, Karuso P. Mechanism of reversible staining of protein with epicocconone. Org Lett 7 (2005) 2401-2404.
3) Tannu NS, Sanchez-Brambila G, Kirby P, Andacht TM. Effect of staining reagent on peptide mass fingerprinting from in-gel trypsin digestions: A comparison of SyproRuby". and DeepPurple".. Electrophoresis 27 (2006) 3136-3143.

Deep purple does not contain any heavy metals and is completely reversible!


Dye Hard

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (luigi.villa)
« Reply #2 on: December 13, 2006, 05:00:00 PM »
For the best cost/quality ratio far beyond Flamingo, Sypro Ruby and Deep Purple check:

Proteomics. 2004 Mar;4(3):599-608.
Improved Ruthenium II tris (bathophenantroline disulfonate) staining and destaining protocol for a better signal-to-background ratio and improved baseline resolution.
Lamanda A, Zahn A, Roder D, Langen H.


Sir Stainalot

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Re: Flamingo, Sypro Ruby and Deep Purple (Dye Hard)
« Reply #3 on: December 13, 2006, 05:00:00 PM »
For the practical use of RuBPS staining check the following References:

Miller, I.; Crawford, J.; Gianazza, E. Proteomics 2006.

Bryborn, M.; Adner, M.; Cardell, L. O. Respir Res 2005, 6, 118.

Clerk, A.; Cullingford, T. E.; Kemp, T. J.; Kennedy, R. A.; Sugden, P. H. Adv Enzyme Regul 2005, 45, 94-111.

Schaller, A.; Troller, R.; Molina, D.; Gallati, S.; Aebi, C.; Stutzmann Meier, P. Proteomics 2006, 6, (1), 172-80.

Stasyk, T.; Morandell, S.; Bakry, R.; Feuerstein, I.; Huck, C. W.; Stecher, G.; Bonn, G. K.; Huber, L. A. Electrophoresis 2005, 26, (14), 2850-4.

Berger, K.; Wissmann, D.; Ihling, C.; Kalkhof, S.; Beck-Sickinger, A.; Sinz, A.; Paschke, R.; Fuhrer, D. Mol Cell Endocrinol 2004, 227, (1-2), 21-30.

Gorg, A.; Weiss, W.; Dunn, M. J. Proteomics 2004, 4, (12), 3665-85.

Smejkal, G. B.; Robinson, M. H.; Lazarev, A. Electrophoresis 2004, 25, (15), 2511-9.

Junca, H.; Plumeier, I.; Hecht, H. J.; Pieper, D. H. Microbiology 2004, 150, (Pt 12), 4181-7.

Tang, H. Y.; Speicher, D. W. Expert Rev Proteomics 2005, 2, (3), 295-306.

Piette, A.; Derouaux, A.; Gerkens, P.; Noens, E. E.; Mazzucchelli, G.; Vion, S.; Koerten, H. K.; Titgemeyer, F.; De Pauw, E.; Leprince, P.; van Wezel, G. P.; Galleni, M.; Rigali, S. J Proteome Res 2005, 4, (5), 1699-708.

Quaglino, D.; Boraldi, F.; Bini, L.; Volpi, N. Current Proteomics 2004, 1, 167-178.

Hjerno, K.; Alm, R.; Canback, B.; Matthiesen, R.; Trajkovski, K.; Bjork, L.; Roepstorff, P.; Emanuelsson, C. Proteomics 2006, 6, (5), 1574-87.

Gerber, I. B.; Laukens, K.; Witters, E.; Dubery, I. A. Plant Physiol Biochem 2006, 44, (5-6), 369-79.

Chevallet, M.; Diemer, H.; Luche, S.; van Dorsselaer, A.; Rabilloud, T.; Leize-Wagner, E. Proteomics 2006, 6, (8), 2350-4.



Michael

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Re: Flamingo, Sypro Ruby and Deep Purple (luigi.villa)
« Reply #4 on: December 13, 2006, 05:00:00 PM »
In our hands, Flamingo has far and away the best signal-to-noise, sensitivity and simplicity.  We don't have a comparison of the MS results (in house, yet), but we aren't worried about it.  Cost is not really a consideration for us as they are all equivalent (in our calculations) and the home-made RuBP is not an option.  As for DIGE utility, we don't do DIGE so can't tell you.

Dances with Dyes

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Michael)
« Reply #5 on: December 13, 2006, 05:00:00 PM »

RuBPS is unbeatable concerning the costs and the broad aplication range of the molecule. It was never "home-made" as it was developped, patent protected and published  in the 80ies by Bannwarth et al. at Hoffmann La-Roche in Basel.  

Bannwarth, W.; Schmidt, D.; Stallard, R. L.; Hornung, C.; Knorr, R.; Müller, F. Helv Chim Acta 1988, 71, 2085-2099.

Since the Rabilloud publication in 2000 there were never ending rumours about the RuBPS of beeing SYPRO Ruby which is simply not true. Just read the SYPRO Patent.

RuBPS is known to be rock stable. It can be stored in dried form for years.

You best read:

Miller et al.
Protein stains for proteomic applications:
Which, when, why? Proteomics 2006, 6 online


Ambort

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Dances with Dyes)
« Reply #6 on: December 14, 2006, 05:00:00 PM »
Ciao Lamanda,

can you not just register as user instead of using all those different guest names?

best regards

Ambort


Ardea

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Ambort)
« Reply #7 on: December 14, 2006, 05:00:00 PM »
Does anyone know if you can buy the RuBP solution ready to use???
Although it looks pretty simple to make from the Rabilloud 2001 publication I would prefer to buy it to avoid making any mistakes.

Buran

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Ardea)
« Reply #8 on: December 14, 2006, 05:00:00 PM »
Just go to Product Number 03038 from Fluka on (page 2019 in the catalogue 07/08) or http://www.sigmaaldrich.com/catalog/search/ProductDetail/FLUKA/03038" TARGET="_blank">http://www.sigmaaldrich.com/ca...03038. The solution is not ready to use, but you only need to dilute it pror to use.

Greetings Jury from Russia

cchan

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Ardea)
« Reply #9 on: December 14, 2006, 05:00:00 PM »
Hi,

you can get RuBPS from Biotium (cat # 80021). I have used this to stain and it works well. It comes in a 5 mg solid package which you just dilute with water to get 20 mM (I think its about 150 ul) stock.

Hope this helps

Chun


Michael

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Re: Flamingo, Sypro Ruby and Deep Purple (Dances with Dyes)
« Reply #10 on: December 14, 2006, 05:00:00 PM »
How does RuBPS compare to Flamingo?  I scanned the review and it doesn't mention Flamingo (that I could see).  At one time, not now I see, you could not buy RuBPS so it wasn't even a consideration.

Dances with Dyes

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Michael)
« Reply #11 on: December 14, 2006, 05:00:00 PM »
We were beta testers for the BioRad Flamingo. But the Flamingo was suffering from avian flu compared to RuBPS. The same was found for Roti Red and Roti Green.

Michael

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Dances with Dyes)
« Reply #12 on: December 14, 2006, 05:00:00 PM »
Please provide a straight answer...

cchan

  • Guest
Re: Flamingo, Sypro Ruby and Deep Purple (Michael)
« Reply #13 on: December 14, 2006, 05:00:00 PM »
Just out of interest, has any one tried the Krypton Protein stain from Pierce? They seem to claim that theirs works comparably to Sypro and Flamingo but at a much lower cost. Only downside I see is that it cannot be excited by UV.


ardea

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Re: Flamingo, Sypro Ruby and Deep Purple (cchan)
« Reply #14 on: December 14, 2006, 05:00:00 PM »
I got a free test sample of Krypton from Pierce and it showed similar sensitivity as my silver stainings. I did not compare it directly to Sypro or any other flourescent dye, so I can not say if it is more or less sensitive. I got a lot of speckles when I scaned my gel on a Typhoon scanner but Pierce claims that they have not encountered this problem before. If that is true I think Krypton is a pretty good choice.