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® 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® 514	1013.95	511	527		75,000	0.83	0.265	0.116
iFluor® 532	914.06	537	560		90,000	0.68	0.26	0.16
iFluor® 546	1145.41	541	557		100,000	0.67	0.25	0.15
iFluor® 555	1125.26	557	570		100,000	0.64	0.23	0.14
iFluor® 560	1257.45	560	571		120,000	0.57	0.0482	0.069
iFluor® 568	1173.46	568	587		100,000	0.57	0.34	0.15
iFluor® 594	1160.42	588	604		180,000	0.53	0.05	0.04
iFluor® Ultra 594	1436.72	586	601		180,000	N/D	N/D	N/D
iFluor® 597	1058.29	598	618		100,000	0.7	N/D	0.514
iFluor® 610	1211.44	610	628		110,000	0.85	0.32	0.49
iFluor® 633	1249.58	640	654		250,000	0.29	0.062	0.044
iFluor® 647	1274.66	656	670		250,000	0.25	0.03	0.03
iFluor® Ultra 647	2634.28	655	670		250,000	N/D	N/D	N/D
iFluor® 660	1281.66	663	678		250,000	0.26	0.07	0.08
iFluor® 665	1210.53	667	692		N/D	N/D	N/D	N/D
iFluor® 670	923.15	671	682		200,000	0.55	0.03	0.033
iFluor® 680	957.17	684	701		220,000	0.23	0.097	0.094
iFluor® 690	1462.75	685	704		220,000	N/D	N/D	N/D
iFluor® 700	977.16	690	713		220,000	0.23	0.09	0.04
iFluor® 710	1206.39	717	739		190,000	N/D	0.12	0.07
iFluor® 720	1089.29	716	740		140,000	0.14	N/D	N/D
iFluor® 740	1551.19	742	764		125,000	N/D	N/D	N/D
iFluor® 750	1416.83	757	779		275,000	0.12	0.044	0.039
iFluor® Ultra 750	1426.78	749	773		250,000	N/D	N/D	N/D
iFluor® 770	1082.27	777	797		250,000	N/D	N/D	N/D
iFluor® A7	1288.58	762	782		275,000	0.1	0.03	0.03
iFluor® 780	1526.90	784	808		250,000	N/D	N/D	N/D
iFluor® 790	1768.30	787	812		250,000	0.13	0.1	0.09
iFluor® 800	1541.91	801	820		250,000	0.11	0.03	0.08
iFluor® 810	1576.03	811	822		250,000	0.05	0.09	0.15
iFluor® 820	1585.92	822	850		250,000	N/D	0.11	0.16
iFluor® 840	1422	836	879		200,000	N/D	0.2	0.09
iFluor® 860	1571.90	853	878		250,000	N/D	0.1	0.14

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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.