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Chromeo™ Py-Dyes are a new class of amine-reactive fluorescent pyrylium dyes that change their color upon reacting with primary amines. They undergo a large shortwave spectral shift (Figure 1), as well as a large increase in quantum yield resulting in a bright fluorescent conjugate (see the Properties tab below). This effectively eliminates background from any unbound dye. In addition, because unbound Py-Dye is hydrolyzed during the labeling procedure, there is no need for purification following the simple one-step, room-temperature incubation. The labeled protein is ready to use immediately, and any possible background from unbound dye is eliminated. Py-Dyes are stable in aqueous solvents and are suitable for a variety of applications, such as labeling proteins or as fluorescent stain of proteins in electrophoresis.
Figure 1: Color shift of Chromeo Py-Dyes caused by dye conjugation.
Chromeo P503 is blue while in its free state. After the short conjugation reaction, the dye's color undergoes a large Stokes shift and becomes red.
| Name | Format | Cat. No. | Price | |
|---|---|---|---|---|
| Chromeo™ P465 | 1 mg | 15105 | $205 |
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| 5 mg | 16105 | $695 |
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| Chromeo™ P503 | 1 mg | 15106 | $205 |
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| 5 mg | 16106 | $695 |
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| Chromeo™ P540 | 1 mg | 15107 | $205 |
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| 5 mg | 16107 | $695 |
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| Chromeo™ P429 | 1 mg | 15108 | $205 |
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| 5 mg | 16108 | $695 |
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| Chromeo™ P543 | 1 mg | 15109 | $205 |
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| 5 mg | 16109 | $695 |
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| Cell Biology & Fluorescence Products Brochure |
| Chromeo™ P429 Data Sheet |
| Chromeo™ P465 Data Sheet |
| Chromeo™ P503 Data Sheet |
| Chromeo™ P540 Data Sheet |
| Chromeo™ P543 Data Sheet |
Chromeo™ Py-Dye advantages
- No washing steps required
- No background of unlabeled dye
- Fast and simple procedure
- No change in net charge of the labeled molecule
- No interference of excitation and emission wavelength
Because of their unique features, the Chromeo Py-Dyes (Chromeo P429, P456, P503, P540 and P543) have been used successfully in a number of applications such capillary electrophoresis, SDS-gel electrophoresis or isoelectric focusing.
Application references for fluorescent Py-Dyes
Chromeo P503 used as a protein stain in SDS-gel electrophoresis
"SDS-PAGE of proteins using a chameleon-type of fluorescence prestain" by Meier RJ, Steiner MS, Duerkop A, Wolfbeis OS. Anal Chem. August 15, 2008. Vol 80(16):6274-9.
Chromeo P503 used to monitor amino-bonds on surfaces
"Monitoring of Amino Functionalities on Plasma-Chemically Modified Polypropylene Supports with a Chromogenic and Fluorogenic Pyrylium Reporter" by Katrin Hoffmann, Renate Mix, Ute Resch-Genger and Joerg F. Friedrich. Langmuir 2007, Vol. 23, 8411-16.
Chromeo P503 and Chromeo P540 used as a protein-stain in capillary electrophoresis (CE)
"Determination of picomolar concentrations of protein using novel amino reactive chameleon labels and capillary electrophoresis laser-induced fluorescence detection" by Douglas B. Craig, Bianca Wetzl, Axel Duerkop and Otto S. Wolfbeis. Electrophoresis. 2005. Vol. 26. 2208-13.
"Reaction of organic reagents with proteins III. Spectroscopic and electrophoretic behavior of proteins labeled with Chromeo P503" by Emily H. Turner, Jane A. Dickerson, Lauren M. Ramsay, Kristian E. Swearingen, Roza Wojcik and Norman J. Dovichi. Journal of Chromatography A. 2008. Vol. 1194. 253-6.
Chromeo P540 used as a protein-stain in capillary isoelectric focusing
"Femtomolar Concentration Detection Limit and Zeptomole Mass Detection Limit for Protein Separation by Capillary Isoelectric Focusing and Laser-Induced Fluorescence Detection" by Lauren M. Ramsay, Jane A. Dickerson, Oluwatosin Dada, and Norman J. Dovichi* Analytical Chemistry, 2009, February 10.
| Dye | Dye State | Absorption | Emission | ε L/(mol-cm) | Quantum Yield* |
|---|---|---|---|---|---|
| Chromeo™ P465 | Free | 645 nm | 732 nm | 25,000 | < 1% |
| Conjugated | 465 nm | 630 nm | 25,000 | ~14% | |
| Chromeo™ P503 | Free | 612 nm | 665 nm | 60,000 | < 1% |
| Conjugated | 503 nm | 600 nm | 24,000 | ~50% | |
| Chromeo™ P540 | Free | 587 nm | – | 80,000 | 0% |
| Conjugated | 533 nm | 627 nm | 50,000 | ~20% | |
| Chromeo™ P429 | Free | 457 nm | – | 65,000 | 0% |
| Conjugated | 429 nm | 536 nm | 75,000 | ~10% | |
| Chromeo™ P543 | Free | 570 nm | – | 110,000 | 0% |
| Conjugated | 543 nm | 590 nm | 57,000 | ~15% | |
| Table 1: Properties of Chromeo Py-Dyes. *Quantum Yield will depend on the DPR of the conjugated protein. |
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Each Chromeo Py-Dye is supplied as a dry powder. Guaranteed stable for 6 months when stored properly.
