Mix-n-Stain™ Fluorescent Protein & Tandem Dye Antibody Labeling Kits allow you to label your antibody with a fluorescent phycobiliprotein in about 4 hours with no purification. The labeling procedure tolerates many common buffer components and antibody stabilizers (see product protocols for details).
- Choice of APC, R-PE (PE), PerCP, or tandem dyes
- Labeling takes a few hours with minimal hands-on time
- Kit sizes for labeling 25-50 ug, 50-100 ug or 1 mg IgG
- No purification, 100% recovery
Fluorescent proteins (APC, R-PE and PerCP):
- APC (Allophycocyanin) is a bright phycobiliprotein of ~105 kDa, found in red algae and cyanobacteria (blue-green algae). It has absorption maxima at 650 and emission at 660 nm. Because of the high fluorescence quantum yield (~0.68), APC is by far the brightest dye for the 650 nm wavelength region.
- R-PE (R-phycoerythrin, also called PE) is a large phycobiliprotein with a molecular weight of ~240 kDa. It has absorption maxima at 496, 546, and 565 nm respectively and emission at 578 nm. Because of the high extinction coefficient (~1,960,000) and high fluorescence quantum yield (~0.82), R-PE is by far the brightest dye for the 565 nm wavelength region. R-PE is one of the brightest fluorochromes for flow cytometry, but due to its photobleaching properties it is not recommended for fluorescence microscopy.
- PerCP (Peridinin Chlorophyll Protein Complex) is a small, 35 kDa, phycobiliprotein found in dinoflagellates. PerCP has absorption maxima at 482 nm and emission at 677 nm.
Tandem dyes are composed of two covalently attached fluorescent molecules. An example of this a phycobiliprotein to which small molecule dyes are conjugated. By a process called Förster resonance energy transfer (FRET), or fluorescence resonance energy transfer, the tandem dye has the excitation properties of the donor molecule and the emission properties of the acceptor. The Stokes shift of the tandem dye is larger than that of the original donor molecule. Tandem dyes are used almost exclusively for flow cytometry because of imperfect energy transfer, and because they show bleed-through fluorescence in the acceptor dye channel, which requires compensation. Aside from cytometers, most fluorescence instruments don’t perform compensation and are generally not configured for the long-stokes shifts of tandem dyes.
We offer kits for labeling with the following tandem dyes featuring Biotium’s next-generation CF® Dyes with superior brightness.
- RPE-CF®583R (Ex/Em 496, 546, 586/609 nm) for the RPE-Texas Red® channel
- RPE-CF®647T (Ex/Em 496, 546, 555/665 nm) for the RPE-Cy®5 channel
- APC-CF®750T (Ex/Em 496, 546, 650/780 nm) for the APC-Cy®7 channel
- APC-CF®790T (Ex/Em 496, 546, 784/806 nm) for the APC-Cy®7 channel.
Note: APC-CF®790T is intended for spectral flow cytometry. For conventional flow cytometry use APC-CF®750T.
We also offer Mix-n-Stain™ Kits for labeling antibodies, nanobodies, or small ligands with bright & photostable CF® dyes and other labels. See our full line of Antibody & Protein Labeling Kits.
Mix-n-Stain™ Fluorescent Protein Antibody Labeling Kits
Note: Kits are not recommended for labeling IgM antibodies. Cy Dye is a registered trademark of GE Healthcare.