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iFluor® Dye Selection Guide
AAT Bioquest iFluor® dyes are a series of highly water-soluble fluorescent dyes for labeling biomolecules, such as proteins and antibodies. Available in a variety of excitation-emission profiles, spanning the UV-visible to near-infrared spectrum, there is an iFluor® dye to match any instrument set-up and many combinations for multiplex detection. Conjugates made with iFluor® dyes exhibit superior brightness and photostability, outperforming most Alexa Fluor® conjugates and other spectrally similar conjugates.
Benefits of iFluor® Dyes and Conjugates
Image of HeLa cells stained with mouse anti-tubulin followed with iFluor® 594 goat anti-mouse IgG. Actin filaments were stained with Phalloidin-iFluor® 488 Conjugate and nuclei were stained with DAPI.
- Brightness: iFluor® conjugates exhibit more intense fluorescence than other spectrally similar conjugates (e.g. Alexa Fluor® dyes, fluorescein dyes, cyanine dyes, etc.)
- Photostability: iFluor® conjugates are more photostable than spectrally similar conjugates allowing for more time to capture images.
- Water solubility: iFluor® reactive dyes are highly water soluble. Conjugation can be performed without organic solvents resulting in conjugates that are resistant to precipitation during storage.
- pH insensitivity: iFluor® dyes are robust and highly fluorescent across a broad pH range.
- Instrument compatibility: Absorption spectra of iFluor® dyes are matched to the principal output wavelengths of common excitation sources.
- Color selection: iFluor® dyes are available in several distinct colors, ranging from violet (350 nm) to infrared (860 nm), maximizing choice and flexibility in multicolor analysis.
iFluor® Dyes Spectral Properties
Table 1. iFluor® Dyes Spectral Properties
| iFluor® Dye ▲ ▼ | Mol. Wt. ▲ ▼ | Abs. (nm) ▲ ▼ | Em. (nm) ▲ ▼ | Spectrum ▲ ▼ | ε¹ ▲ ▼ | Φ² ▲ ▼ | CF at 260 nm³ ▲ ▼ | CF at 280 nm⁴ ▲ ▼ |
| iFluor® 350 | 749.85 | 345 | 450 |  | 20,000 | 0.95 | 0.83 | 0.23 |
| iFluor® 405 | 755.58 | 403 | 427 | | 37,000 | 0.91 | 0.48 | 0.77 |
| iFluor® 430 | 688.79 | 433 | 498 | | 40,000 | 0.78 | 0.68 | 0.3 |
| iFluor® 440 | 692.83 | 434 | 480 | | 40,000 | N/D | 0.352 | 0.229 |
| iFluor® 445 | 968.01 | 446 | 558 | | N/D | N/D | N/D | N/D |
| iFluor® 450 | 603.69 | 451 | 502 | | 40,000 | 0.82 | 0.45 | 0.27 |
| iFluor® 460 | 793.99 | 468 | 493 | | 80,000 | 0.8 | 0.98 | 0.46 |
| iFluor® 488 | 945.07 | 491 | 516 | | 75,000 | 0.9 | 0.21 | 0.11 |
| iFluor® 500 | 981.12 | 501 | 520 | | N/D | N/D | N/D | N/D |
| iFluor® 510 | 951.91 | 511 | 530 | | N/D | N/D | N/D | N/D |
- Ext. Coeff. = Extinction coefficient at their maximum absorption wavelength. The units of extinction coefficient are cm-1M-1.
- FQY = fluorescence quantum yield in aqueous buffer (pH 7.2).
- CF at 260 nm is the correction factor used for eliminating the dye contribution to the absorbance at 260 nm.
- CF at 280 nmis the correction factor used for eliminating the dye contribution to the absorbance at 280 nm (for peptides and protein labeling).
- Fluorescence intensity is significantly increased upon coupling to proteins.
iFluor® Dye Selection Guide
| Fluorophore | Ex/Em (nm) | Alternative to | Reative Targets | ||||||
| Amine | Thiol | Aldehyde | Alkyne | Azide | |||||
| iFluor® 350 | 344/488 | Coumarin • Alexa Fluor® 350 • DyLight™ 350 | #1020 | #1060 | #1080 | #1070 | |||
| iFluor® 405 | 402/425 | Cascade Blue® • Alexa Fluor® 405 • DyLight™ 405 | #1021 | #1053 | #1081 | #1071 | |||
| iFluor® 430 | 433/495 | Alexa Fluor® 430 | #1052 | #1054 | |||||
| iFluor® 440 | 434/480 | #1041 | |||||||
| iFluor® 450 | 451/502 | #1026 | #1057 | ||||||
| iFluor® 488 | 492/516 | Alexa Fluor® 488 • DyLight™ 488 | #1023 | #1062 | #1082 | #1072 | #1000 | #999 | |
| iFluor® 514 | 527/554 | Alexa Fluor® 514 | #1024 | ||||||
| iFluor® 532 | 543/563 | Alexa Fluor® 532 | #1025 | #1061 | |||||
| iFluor® 546 | 541/557 | Alexa Fluor® 546 | #1048 | ||||||
| iFluor® 555 | 556/569 | Cy3® • Alexa Fluor® 555 • DyLight™ 550 | #1028 | #1063 | #1083 | #1073 | #1093 | #1092 | |
| iFluor® 560 | 559/571 | #1040 | |||||||
| iFluor® 568 | 568/587 | Rhodamine Red • Alexa Fluor® 568 | #1049 | #1055 | |||||
| iFluor® 594 | 587/603 | Texas Red® • Alexa Fluor® 594 • DyLight™ 594 | #1029 | #1064 | |||||
| iFluor® 610 | 609/627 | Alexa Fluor® 610 | #1038 | ||||||
| iFluor® 633 | 638/652 | Alexa Fluor® 633 | #1030 | #1056 | |||||
| iFluor® 647 | 654/669 | Cy5® • Alexa Fluor® 647 • DyLight™ 650 | #1031 | #1065 | #1085 | #1074 | #1091 | #1090 | |
| iFluor® 660 | 660/677 | Alexa Fluor® 660 | #1032 | #1075 | |||||
| iFluor® 670 | 669/682 | Cy5® B | #1033 | ||||||
| iFluor® 680 | 683/700 | Cy5.5® • IRDye® 700 • Alexa Fluor® 680 | #1035 | #1066 | #1086 | #1076 | |||
| iFluor® 700 | 690/713 | Alexa Fluor® 700 | #1036 | #1067 | #1087 | #1077 | |||
| iFluor® 710 | 712/736 | #1045 | #1078 | ||||||
| iFluor® 750 | 759/777 | Cy7® • Alexa Fluor® 750 • DyLight™ 750 | #1037 | #1068 | #1088 | #1079 | |||
| iFluor® 790 | 786/811 | IRDye® 800 • Alexa Fluor® 790 • DyLight™ 800 | #1368 | #1360 | #1366 | #1364 | #1362 | ||
| iFluor® 800 | 801/820 | #1379 | #1375 | #1378 | |||||
| iFluor® 810 | 812 | #1389 | #1385 | #1388 | |||||
| iFluor® 820 | 820 | #1399 | #1395 | #1398 | |||||
| iFluor® 840 | 836 | #1403 | #1401 | #1402 | |||||
| iFluor® 860 | 854 | #1409 | #1405 | #1408 | |||||
| Succinimidyl Ester | Acid | Maleimide | Hydrazide | Amine | Azide | Alkyne | |||
| Reactive Forms | |||||||||
Original created on August 28, 2019, last updated on November 8, 2022
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