148_12_AKR Fluorochromes Poster02:Layout 1 23/04/2012 15:10 Page 1

A Guide to
Fluorochromes

Excitation and Emission Spectral Profiles Dye Max. excitation

wavelength (nm)
Max.emission
wavelength (nm)
Excitation laser
lines (nm)
Methoxycoumarin 360 410
Relative Intensity

Laser line (488)

Excitation Single DyLight® 405
Alexa Fluor® 405
400
402
420
421
Emission PE
(PE) Brilliant Violet 421™
HiLyte FluorTM 405
407
404
421
428
DyLight® 350 353 432
Alexa Fluor® 350 346 442
Aminocoumarin (AMCA) 350 445
BD Horizon™ V450 404 448
Pacific BlueTM 404 456 360,405,407
350 400 450 500 550 600 650 700 750 800 EviTagTM quantum dots-Lake Placid Blue 470 490
AMCyan 457 491
Wavelength (nanometers) BD Horizon™ V500 415 500
Cy2® 489 506 488
ChromeoTM 488 488 517
Relative Intensity

Laser line (633) DyLight® 488 493 518

Excitation Single Alexa Fluor® 488 495 519 488
Emission Cy5® FAM 494 519
(Cy5®) Fluorescein Iso-thiocyanate (FITC) 495 519 488
EviTagTM quantum dots-Adirondack Green 505 520
ChromeoTM 505 505 526
HiLyte FluorTM 488 501 527
Alexa Fluor® 514 518 540
EviTagTM quantum dots-Catskill Green 525 540
Alexa Fluor® 430 434 541
350 400 450 500 550 600 650 700 750 800 Pacific Orange™ 403 551
Wavelength (nanometers) Alexa Fluor® 532 532 554
HEX 535 556
EviTagTM quantum dots-Hops Yellow 545 560
Relative Intensity

Laser line (488) ChromeoTM 546 545 561

Excitation Tandem Cy3® 548 561 488,514
Emission PE-Cy5® Alexa Fluor® 555 555 565
(PE-Cy5®) HiLyte FluorTM 555 550 566
5-TAMRA 541 568
Alexa Fluor® 546 556 573 532
DyLight® 550 562 576
Phycoerythrin (PE) 496,566 576 488
Tetramethyl Rhodamine Isothiocyanate (TRITC) 557 576
EviTagTM quantum dots-Birch Yellow 560 580
350 400 450 500 550 600 650 700 750 800 Cy3.5® 576 589 568,543
Rhodamine Red-X 570 590
Wavelength (nanometers)
PE-Dyomics® 590 488 599
Fluorochromes have unique and characteristic spectra for absorption (excitation) and emission. A single dye EviTagTM quantum dots-Fort Orange 585 600
is excited at a particular wavelength and emits a photon at a higher wavelength. A tandem dye consists of a ROX 575 602
donor and acceptor fluorochrome molecule, placed in close proximity, allowing for energy transfer between Alexa Fluor® 568 578 603 532
the two. The tandem dye is excited at the excitation wavelength of the donor molecule and emits a photon at Red 613 480,565 613
the emission wavelength of the acceptor molecule. Texas Red® 595 613 568,543,514
HiLyte FluorTM 594 593 616
PE-Texas Red® 566 616
Energy Level Diagrams Alexa Fluor® 594
DyLight® 594
590
593
617
618
EviTagTM quantum dots-Maple-Red Orange 600 620
Alexa Fluor® 610 612 628
Single dye ChromeoTM 494 494 628
Alexa Fluor® 633 632 647
SureLight® APC 652 657
DyLight® 633 638 658
Allophycocyanin (APC) 650 660 595,633,635,647
Es ChromeoTM 642 642 660
Fluorescence Quantum Red 488 660
Excitation 2 3 SureLight® P3 614 662
(Emission)
Alexa Fluor® 647 650 665 595,633,635,647
Cy5® 647 665 633,635
PE-Cy5® 565 666 488
Laser 1 Gs SureLight® P1 545 666
PE-Alexa Fluor® 647 567 669
PE-Dyomics® 647 488 672
Tandem dye DyLight® 650 654 673
HiLyte FluorTM 647 650 675
Peridinin Chlorophyll (PerCP) 477 678 488
IRDye® 700DX 680 687
Alexa Fluor® 660 663 690
Es PE-Cy5.5® 565 693 488
APC-Cy5.5® 650 694 595,633,635,647
Cy5.5® 675 694 647
D 3 A Fluorescence TruRed 490,675 695
Excitation 2 4 (Emission) HiLyte FluorTM 680 678 699
Alexa Fluor® 680 679 702
DyLight® 680 692 712
Alexa Fluor® 700 702 723
APC-Cy7® 650 774 595,633,635,647
Laser 1 Gs Alexa Fluor® 750 749 775
Cy7® 753 775
PE-Dyomics® 747 488 776
DyLight® 755 754 776
For a single fluorescent dye, a laser set at the signature excitation wavelength for the dye provides HiLyte FluorTM 750 753 778
electromagnetic energy to an electron in that molecule (1).1 The electron moves to an excitation state at the PE-Cy7® 566 778 488
next energy level (Es) (2).
2 Energy is then released in the form of a photon (fluorescence) and the electron IRDye® 800RS 770 786
moves back down to the lower energy level (Gs) (3). 3 In the case of a tandem fluorescent dye, after excitation DyLight® 800 777 794
of the electron by a laser (11 - 2),
2 energy is released by an electron in the donor molecule (D)
D and absorbed IRDye® 800CW 778 794
by an electron in the acceptor molecule (A) (4). The electron in the acceptor molecule moves to an excitation
A 3 Alexa Fluor® 790 782 805
state at the next energy level (Es). Similar to a single dye, energy is then released in the form of the photon
(fluorescence) and the electron moves back down to the lower energy level (Gs) (5). 4

Nucleic Acid Dyes Max. excitation Max. emission Excitation laser

Multicolor staining example wavelength (nm) wavelength (nm) lines (nm)
DAPI 359 461 325,360,405,407
MERGED ab16051 - Beta Catenin DRAQ5™ (ab108410) allows rapid staining of Hoechst 33258 352 461
dsDNA/nuclei of LIVE or FIXED cells. It is Hoechst 33342 350 461
ideal for use in multi-color analysis and SYTOX Blue 431 480
compatible with GFP labels. As well as
YOYO-1 491 509
offering half the scan times of Hoechst or
SYTOX Green 504 523
DAPI, DRAQ5™ can be used in a greater
variety of applications. The image on the left
TOTO-1, TO-PRO-1 509 533
shows HeLa cells stained with 5μM DRAQ5™ Mithramycin 450 570
DRAQ5 ab7291 - Alpha Tubulin
to label nuclear DNA (pseudocolor orange). SYTOX Orange 547 570
Pre-absorbed goat polyclonal secondary Chromomycin A3 445 575
antibodies to rabbit IgG - H&L were used to CyTRAK Orange™* 457,488,549 615
visualise beta Catenin (DyLight® 488, Ethidium Bromide 493 620
ab96899, green) and alpha Tubulin (DyLight® Propidium iodide (PI) 305,540 620 325,360,488
594, ab96899, red). DRAQ5™ 646 681,697
DRAQ7™ 599, 644 678,697
148_12_AKR

*CyTRAK Orange™ stains both nucleus and cytoplasm

Discover more at abcam.com/fluorochromes

Copyright © 2012 Abcam, All Rights Reserved. The Abcam logo is a registered trademark. All information/detail is correct at time of going to print. Color bars are an approximation of corresponding emission wavelength.
Dylight® is a trademark of Thermo Fisher Scientific and its subsidiaries. Cy® and CyDye® are trademarks of GE Healthcare Limited. Alexa Fluor® is a trademark of Molecular Probes Inc. HILYTE FLUOR™ is a trademark of AnaSpec, Inc. DRAQ5™, DRAQ7™ and CyTRAK Orange™are trademarks of Biostatus Ltd.