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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® 350749.8534545020,0000.950.830.23
iFluor® 405755.5840342737,0000.910.480.77
iFluor® 430688.7943349840,0000.780.680.3
iFluor® 440692.8343448040,000N/D0.3520.229
iFluor® 450603.6945150240,0000.820.450.27
iFluor® 460793.9946849380,0000.80.980.46
iFluor® 488945.0749151675,0000.90.210.11
iFluor® 5141013.9551152775,0000.830.2650.116
iFluor® 532914.0653756090,0000.680.260.16
iFluor® 5461145.41541557100,0000.670.250.15
  1. Ext. Coeff. = Extinction coefficient at their maximum absorption wavelength. The units of extinction coefficient are cm-1M-1.
  2. FQY = fluorescence quantum yield in aqueous buffer (pH 7.2).
  3. CF at 260 nm is the correction factor used for eliminating the dye contribution to the absorbance at 260 nm.
  4. CF at 280 nmis the correction factor used for eliminating the dye contribution to the absorbance at 280 nm (for peptides and protein labeling).
  5. 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 August 28, 2019
Tagged under: Antibody & Protein Labeling