iFluor Dyes and Kits

AAT Bioquest iFluor™ dyes are a series of highly water-soluble fluorescent dyes for covalent labeling of 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 Alexa Fluor® conjugates and other spectrally similar conjugates. Use iFluor™ products for all your immunofluorescence assays, cell imaging and flow cytometric applications.

iFluor™ Reactive Dyes


iFluor™ reactive dyes may be covalently labeled to biomolecules without self-quenching, producing intensely fluorescent conjugates. iFluor™ reactive dye formats include amine-reactive succinimidyl ester, thiol-reactive maleimide, and more. For assistance labeling proteins with succinimidyl ester or maleimide iFluor™ dyes, use our online interactive protein labeling protocol.

Available Labeling Chemistries

  • Succinimidyl Esters: for labeling primary amines (-NH2) on proteins, antibodies, peptides, nucleic acids and other biomolecules. They are commonly used to prepare bioconjugates for immunochemistry, fluorescence in situ hybridization (FISH), cell tracing, receptor labeling and fluorescent analog cytochemistry.
  • Maleimides: for labeling thiols or sulfhydryl groups (-SH) to selectively modify a protein at a defined site. Thiol-reactive dyes are often used to prepare fluorescent peptides, proteins and oligonucleotides for probing biological structures, functions and interactions.
  • Azides: for labeling ethylene groups via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction.
  • Alkynes: for labeling azide groups via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction.
  • Carboxylic acids: for labeling amines after pre-activation with carbodiimides or for Steglich esterification of alcohols.
  • Amines: for labeling various electrophilic compounds such as activated esters.

The table below shows the range of available iFluor™ reactive dyes, their excitation and emission maxima, and their fluorophore equivalents to assist you in selecting the best fluorophore for labeling proteins, antibodies, and other biopolymers, as well as, derivatizing low molecular weight molecules.

 

Table 1. Available iFluor™ Reactive Dyes for Labeling Biomolecules.

Fluorophore Ex (nm) Em (nm) Alternative to Amine Reactive Thiol Reactive Alkyne Reactive Azide Reactive Aldehyde/ Ketone Reactive Carbonyl-Reactive Click- Chemistry
iFluor™ 350 344 448 Coumarin • Alexa Fluor® 350 iFluor™ 350 succinimidyl ester iFluor™ 350 maleimide iFluor™ 350 hydrazide iFluor™ 350 amine
iFluor™ 405 402 425 Cascade Blue® • Alexa Fluor® 405 iFluor™ 405 succinimidyl ester iFluor™ 405 maleimide iFluor™ 405 hydrazide iFluor™ 405 amine
iFluor™ 430 433 495 Alexa Fluor® 430 iFluor™ 430 succinimidyl ester iFluor™ 430 maleimide
iFluor™ 450 451 502 iFluor™ 450 succinimidyl ester iFluor™ 450 maleimide
iFluor™ 488 491 516 FITC • Alexa Fluor® 488 iFluor™ 488 succinimidyl ester iFluor™ 488 maleimide iFluor™ 488 azide iFluor™ 488 alkyne iFluor™ 488 hydrazide iFluor™ 488 amine iFluor™ 488 Tetrazine

iFluor™ 488 TCO

iFluor™ 514 527 554 Alexa Fluor® 514 iFluor™ 514 succinimidyl ester
iFluor™ 532 543 563 Alexa Fluor® 532 iFluor™ 532 succinimidyl ester iFluor™ 532 maleimide
iFluor™ 546 541 557 Alexa Fluor® 546 iFluor™ 546 succinimidyl ester
iFluor™ 555 556 569 Cy3® • Alexa Fluor® 555 iFluor™ 555 succinimidyl ester iFluor™ 555 malemide iFluor™ 555 azide iFluor™ 555 alkyne iFluor™ 555 hydrazide iFluor™ 555 amine iFluor™ 555 Tetrazine

iFluor™ 555 TCO

iFluor™ 560 559 571 iFluor™ 560 succinimidyl ester
iFluor™ 568 568 587 Rhodamine Red • Alexa Fluor® 568 iFluor™ 568 succinimidyl ester iFluor™ 568 maleimide
iFluor™ 594 587 603 Texas Red® • Alexa Fluor® 594 iFluor™ 594 succinimidyl ester iFluor™ 594 maleimide iFluor™ 594 Tetrazine

iFluor™ 594 TCO

iFluor™ 610 609 627 Alexa Fluor® 610 iFluor™ 610 succinimidyl ester
iFluor™ 633 638 652 Alexa Fluor® 633 iFluor™ 633 succinimidyl ester iFluor™ 633 maleimide
iFluor™ 647 654 669 Cy5® • Alexa Fluor® 647 iFluor™ 647 succinimidyl ester iFluor™ 647 maleimide iFluor™ 647 azide iFluor™ 647 alkyne iFluor™ 647 hydrazide iFluor™ 647 amine iFluor™ 647 Tetrazine

iFluor™ 647 TCO

iFluor™ 660 660 677 Alexa Fluor® 660 iFluor™ 660 succinimidyl ester iFluor™ 660 amine
iFluor™ 670 669 682 Cy5® B iFluor™ 670 succinimidyl ester
iFluor™ 680 683 700 Cy5.5® • IRDye® 700 • Alexa Fluor® 680 iFluor™ 680 succinimidyl ester iFluor™ 680 maleimide iFluor™ 680 hydrazide iFluor™ 680 amine
iFluor™ 700 690 713 Alexa Fluor® 700 iFluor™ 700 succinimidyl ester iFluor™ 700 maleimide iFluor™ 700 hydrazide iFluor™ 700 amine
iFluor™ 710 712 736 iFluor™ 710 succinimidyl ester iFluor™ 710 amine
iFluor™ 750 759 777 Cy7® • Alexa Fluor® 750 iFluor™ 750 succinimidyl ester iFluor™ 750 maleimide iFluor™ 750 hydrazide iFluor™ 750 amine
iFluor™ 790 786 811 IRDye® 800 • Alexa Fluor® 790 iFluor™ 790 succinimidyl ester iFluor™ 790 maleimide iFluor™ 790 hydrazide iFluor™ 790 amine
iFluor™ 800 801 820 iFluor™ 800 succinimidyl ester iFluor™ 800 maleimide
iFluor™ 810 811 822 iFluor™ 810 succinimidyl ester iFluor™ 810 maleimide
iFluor™ 820 820 849 iFluor™ 820 succinimidyl ester iFluor™ 820 maleimide
iFluor™ 840 836 876 iFluor™ 840 succinimidyl ester iFluor™ 840 maleimide
iFluor™ 860 852 877 iFluor™ 860 succinimidyl ester iFluor™ 860 maleimide

ReadiLink™ Rapid and xtra Rapid iFluor™ Antibody Labeling Kits


ReadiLink™ Rapid iFluor™ Antibody Labeling Kits and BSA-compatible ReadiLink™ xtra Rapid iFluor™ Antibody Labeling Kits provide a quick and convenient method to label microscale volumes of antibodies with our superior iFluor™ dyes. The unique chemistry of ReadiLink™ kits enable researchers to effortlessly label and recover 100% of their antibodies without a purification step. Since ReadiLink™ conjugates are covalently labeled they are stable for long-term storage and they are ideal for demanding applications including fluorescent microscopy, multi-color imaging, multiplex flow cytometry, primary detection, IHC, FISH, ELISAs and more. Also available are ReadiLink™ Rapid and xtra Rapid Antibody Labeling kits for conjugation of mFluor™ dyes, Alexa Fluor&reg dyes, other fluorescent dyes, biotin, BSA and KLH.

Key features of ReadiLink™ Rapid Antibody Labeling Kits

  • ReadiLink™ xtra Rapid iFluor™ Antibody Labeling Kits are compatible with BSA and other stabilizing proteins
  • Label monoclonal antibodies, polyclonal antibodies or other proteins
  • Quick and simple two-step labeling protocol, no chemistry or conjugation experience necessary
  • Useful for labeling 100 µg of antibody (two separate labeling reactions of 50 µg antibody)
  • 100% antibody recovery, NO purification of the product is required
ReadiLinkWorkflow
Figure 1. ReadiLink™ Rapid Antibody Labeling Kit workflow (figure drawn in BioRender).
ReadiLink™ Antibody Labeling Kit Fluorophore Equivalent Ex/Em (nm) Imaging Flow Cytometry Cat No.
ReadiLink™ Rapid iFluor™ 350 AMCA, Alexa Fluor® 350 344/448 Yes 1220
ReadiLink™ xtra Rapid iFluor™ 350 AMCA, Alexa Fluor® 350 344/448 Yes 1950
ReadiLink™ Rapid iFluor™ 488 FITC, Alexa Fluor® 488 491/516 Yes Yes 1255
ReadiLink™ xtra Rapid iFluor™ 488 FITC, Alexa Fluor® 488 491/516 Yes Yes 1955
ReadiLink™ Rapid iFluor™ 555 Cy3, Alexa Fluor® 555 556/569 Yes 1227
ReadiLink™ xtra Rapid iFluor™ 555 Cy3, Alexa Fluor® 555 556/569 Yes 1958
ReadiLink™ Rapid iFluor™ 594 Texas Red®, Alexa Fluor® 594 587/603 Yes 1230
ReadiLink™ xtra Rapid iFluor™ 594 Texas Red®, Alexa Fluor® 594 587/603 Yes 1960
ReadiLink™ Rapid iFluor™ 633 Cy5, Alexa Fluor® 633 638/655 Yes Yes 1260
ReadiLink™ Rapid iFluor™ 647 Cy5, Alexa Fluor® 647 654/669 Yes Yes 1235
ReadiLink™ xtra Rapid iFluor™ 647 Cy5, Alexa Fluor® 647 654/669 Yes Yes 1963
ReadiLink™ Rapid iFluor™ 680 Cy5.5, Alexa Fluor® 680, IRDye® 700 682/701 Yes Yes 1240
ReadiLink™ Rapid iFluor™ 700 Alexa Fluor® 700 693/713 Yes Yes 1245
ReadiLink™ Rapid iFluor™ 750 Cy7, Alexa Fluor® 750 759/777 Yes 1250
ReadiLink™ xtra Rapid iFluor™ 750 Cy7, Alexa Fluor® 750 759/777 Yes 1965
ReadiLink™ Rapid iFluor™ 790 IRDye® 800, Alexa Fluor® 790 782/811 Yes Yes 1265

iFluor™ Secondary Reagents


iFluor™ secondary detection reagents are optimized for use in a wide range of fluorescence-based applications, including cell-imaging, flow cytometry, and Western blotting. iFluor™ conjugates are produced with the highest quality secondary antibodies and streptavidins, and subjected to rigorous quality control assays. To ensure customers receive products with the greatest sensitivity and selectivity, iFluor™ secondary antibodies have been affinity purified and cross-adsorbed providing higher signal to noise ratios with minimal cross-reactivity.

 

iFluor™ Secondary Antibodies

Secondary antibodies conjugated to bright and photostable iFluor™ dyes can be used to detect and visualize low, medium and high-abundance targets in a variety of cell and protein analysis studies. iFluor™ secondary antibodies recognizes IgG heavy chains as well as immunoglobulin light chains (e.g. kappa or lambda) from either human, mouse or rabbit. The advantages of associated with iFluor™ secondary antibody usage includes excellent water solubility, higher signal-to-noise ratios, enhanced sensivitiy, multiplexing capabilities, and pH-insensitive fluorescence over a wide molar range.

Photostability Comparison

Figure 2. Comparison of photobleaching rates of iFluor™ 488 goat anti-mouse IgG (Cat No. 16735) versus FITC goat anti-mouse IgG (Cat No. 16852). Images were taken at 120-second intervals on a Keyence X710 fluorescence microscope. TFI represents total fluorescence intensity as calculated by ImageJ software.

Table 3. Available iFluor™ secondary antibody conjugates.

Label Abs (nm) Ex (nm) Filter Set Antibody Host Reactivity Cat No.
iFluor™ 350 344 448 DAPI Anti-Human IgG (H&L) Goat Human 50041
iFluor™ 405 402 425 DAPI Anti-Human IgG (H&L) Goat Human 50045
iFluor™ 430 433 495 FITC Anti-Human IgG (H&L) Goat Human 50049
iFluor™ 450 451 502 FITC Anti-Human IgG (H&L) Goat Human 50053
iFluor™ 488 491 516 FITC Anti-Human IgG (H&L) Goat Human 50057
iFluor™ 514 527 554 TRITC Anti-Human IgG (H&L) Goat Human 50061
iFluor™ 532 543 563 TRITC Anti-Human IgG (H&L) Goat Human 50065
iFluor™ 546 541 557 TRITC Anti-Human IgG (H&L) Goat Human 50069
iFluor™ 555 556 569 TRITC Anti-Human IgG (H&L) Goat Human 50073
iFluor™ 560 559 571 TRITC Anti-Human IgG (H&L) Goat Human 50077
iFluor™ 568 568 587 Cy3/TRITC Anti-Human IgG (H&L) Goat Human 50081
iFluor™ 594 587 603 Texas Red Anti-Human IgG (H&L) Goat Human 50085
iFluor™ 610 609 627 Texas Red Anti-Human IgG (H&L) Goat Human 50089
iFluor™ 633 638 652 Texas Red Anti-Human IgG (H&L) Goat Human 50093
iFluor™ 647 654 669 Cy5 Anti-Human IgG (H&L) Goat Human 50097
iFluor™ 660 660 677 Cy5 Anti-Human IgG (H&L) Goat Human 50101
iFluor™ 680 683 700 Cy5.5 Anti-Human IgG (H&L) Goat Human 50109
iFluor™ 700 690 713 Cy5.5 Anti-Human IgG (H&L) Goat Human 50113
iFluor™ 710 712 736 Cy5.5 Anti-Human IgG (H&L) Goat Human 50117
iFluor™ 750 759 777 Cy7 Anti-Human IgG (H&L) Goat Human 50121
iFluor™ 790 786 811 Cy7 Anti-Human IgG (H&L) Goat Human 50125
iFluor™ 800 801 820 Cy7 Anti-Human IgG (H&L) Goat Human 50129
iFluor™ 810 811 822 Cy7 Anti-Human IgG (H&L) Goat Human 50133
iFluor™ 820 820 849 Cy7 Anti-Human IgG (H&L) Goat Human 50137
iFluor™ 840 836 876 Cy7 Anti-Human IgG (H&L) Goat Human 50141
iFluor™ 860 852 877 Cy7 Anti-Human IgG (H&L) Goat Human 50145
iFluor™ 350 344 448 DAPI Anti-Mouse IgG (H&L) Goat Mouse 16730
iFluor™ 405 402 425 DAPI Anti-Mouse IgG (H&L) Goat Mouse 16731
iFluor™ 488 491 516 FITC Anti-Mouse IgG (H&L) Goat Mouse 16735
iFluor™ 514 527 554 TRITC Anti-Mouse IgG (H&L) Goat Mouse 16736
iFluor™ 532 543 563 TRITC Anti-Mouse IgG (H&L) Goat Mouse 16737
iFluor™ 546 541 557 TRITC Anti-Mouse IgG (H&L) Goat Mouse 16457
iFluor™ 555 556 569 TRITC Anti-Mouse IgG (H&L) Goat Mouse 16739
iFluor™ 568 568 587 Cy3/TRITC Anti-Mouse IgG (H&L) Goat Mouse 16470
iFluor™ 594 587 603 Texas Red Anti-Mouse IgG (H&L) Goat Mouse 16741
iFluor™ 633 638 652 Texas Red Anti-Mouse IgG (H&L) Goat Mouse 16743
iFluor™ 647 654 669 Cy5 Anti-Mouse IgG (H&L) Goat Mouse 16744
iFluor™ 680 683 700 Cy5.5 Anti-Mouse IgG (H&L) Goat Mouse 16745
iFluor™ 700 690 713 Cy5.5 Anti-Mouse IgG (H&L) Goat Mouse 16746
iFluor™ 750 759 777 Cy7 Anti-Mouse IgG (H&L) Goat Mouse 16748
iFluor™ 790 786 811 Cy7 Anti-Mouse IgG (H&L) Goat Mouse 16750
iFluor™ 350 344 448 DAPI Anti-Rabbit IgG (H&L) Goat Rabbit 16795
iFluor™ 405 402 425 DAPI Anti-Rabbit IgG (H&L) Goat Rabbit 16796
iFluor™ 488 491 516 FITC Anti-Rabbit IgG (H&L) Goat Rabbit 16773
iFluor™ 514 527 554 TRITC Anti-Rabbit IgG (H&L) Goat Rabbit 16801
iFluor™ 532 543 563 TRITC Anti-Rabbit IgG (H&L) Goat Rabbit 16802
iFluor™ 546 541 557 TRITC Anti-Rabbit IgG (H&L) Goat Rabbit 16618
iFluor™ 555 556 569 TRITC Anti-Rabbit IgG (H&L) Goat Rabbit 16803
iFluor™ 568 568 587 Cy3/TRITC Anti-Rabbit IgG (H&L) Goat Rabbit 16804
iFluor™ 594 587 603 Texas Red Anti-Rabbit IgG (H&L) Goat Rabbit 16806
iFluor™ 633 638 652 Texas Red Anti-Rabbit IgG (H&L) Goat Rabbit 16808
iFluor™ 647 654 669 Cy5 Anti-Rabbit IgG (H&L) Goat Rabbit 16809
iFluor™ 680 683 700 Cy5.5 Anti-Rabbit IgG (H&L) Goat Rabbit 16810
iFluor™ 700 690 713 Cy5.5 Anti-Rabbit IgG (H&L) Goat Rabbit 16811
iFluor™ 750 759 777 Cy7 Anti-Rabbit IgG (H&L) Goat Rabbit 16813
iFluor™ 790 786 811 Cy7 Anti-Rabbit IgG (H&L) Goat Rabbit 16815

iFluor™ Styramide™ Signal Amplifying Reagents

Similar to tyramide signal amplification (TSA), Power Styramide™ Signal Amplification (PSA™) uses the catalytic activity of horseradish peroxidase (HRP) for the covalent deposition and binding of labeled-Styramide™ onto a target protein or nucleic acid sequence in situ. Since the added labeled-Styramide™ are deposited in close proximity of the HRP-target site, there is minimal diffusion-related loss of resolution. Moreover, PSA™ imaging technology can be readily added to any application that allows for the integration of HRP into its protocol, such applications include, IHC, ICC, IF, in situ hybridization and ELISA.

Key features of iFluor™ Styramide™ Reagents

  • Increased detection sensitivity more than 100-fold greater than standard IHC, ICC, IF methods
  • Improved fluorescence signal 10 to 50-fold greater than tyramide (TSA) reagents
  • Higher reactivity of iFluor™ Styramide™ radicals allow for a faster, efficient and more robust labeling of styramide conjugates at the HRP-target interaction site
  • Conserve precious antibodies, iFluor™ Styramide™ labeling allows use of significantly less primary antibodies or probe without any sacrifice in detection sensitivity
  • Available as PSA™ imaging kits with poly-HRP, easy-to-use with sufficient reagents for 100 tests

PSA mulitcolor tissue stain

Figure 3. Multiplex Styramide™ labeling of FFPE tissue. Tissue were incubated with rabbit anti-EpCAM antibodies and mouse anti-pan-Keratin antibodies and then labeled with reagents in the iFluor&trade 488 PSA™ Imaging Kit with Goat Anti-Rabbit IgG (Cat No. 45205) and iFluor&trade 555 PSA™ Imaging Kit with Goat Anti-Mouse IgG (Cat No. 45220), respectively. Nuclei were labeled with DAPI (Cat No. 17507).

The table below shows the range of available iFluor™ Styramide™ reagents, their excitation and emission maximas, extinction coefficient and fluorescence quantum yield to assist you in selecting the best labeling reagent for your fluorescence-based imaging.

 

Table 4. iFluor™ Styramide™ Reagents For iFluor™ Secondary Reagents

iFluor™ Styramide™ Ex/Em (nm) Filter Set Ext. Coeff.1 FQY2 Unit Size
iFluor™ 350 Styramide™ 345/442 DAPI 20,000 0.95 100 Test
iFluor™ 488 Styramide™ 491/514 FITC 75,000 0.9 100 Test
iFluor™ 546 Styramide™ 541/557 Cy3/TRITC 100,000 0.67 100 Test
iFluor™ 555 Styramide™ 552/567 Cy3/TRITC 100,000 0.64 100 Test
iFluor™ 568 Styramide™ 568/587 Cy3/TRITC 100,000 0.57 100 Test
iFluor™ 594 Styramide™ 587/603 Cy3/TRITC 180,000 0.53 100 Test
iFluor™ 647 Styramide™ 654/669 Cy5 250,000 0.25 100 Test
iFluor™ 680 Styramide™ 683/700 Cy5 220,000 0.23 100 Test
iFluor™ 700 Styramide™ 690/713 Cy7 220,000 0.23 100 Test
iFluor™ 750 Styramide™ 759/777 Cy7 275,000 0.12 100 Test
iFluor™ 790 Styramide™ 786/811 Cy7 250,000 0.13 100 Test
  1. Ext. Coeff. = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1).
  2. FQY = fluorescence quantum yield in aqueous buffer (pH 7.2).

iFluor™ Bioconjugates


We offer a large selection of iFluor™-labeled conjugates for a wide range of applications. Featured conjugates include, Annexin V for apoptic detection and phalloidin for cell structure labeling.

 

Annexin V iFluor™ Conjugates for Apoptosis Detection

Annexin V conjugated to iFluor™ dyes provide a fast and reliable method for detecting externalized PS, a key characteristic in early-stage apoptosis. The superior brightness and photostability of Annexin V iFluor™ conjugates are useful for live cell imaging, immunofluorescence, and flow cytometry, and can be combined with other dyes, such as nucleic acid stains, to accurately assess mixed populations of apoptotic, necortic and dead cells.

Key features of Annexin V iFluor™ Conjugates

  • iFluor™ conjugates generate brighter more photostable signals
  • Compatible with live cells only
  • Fixable after staining with formaldehyde
  • Suitable for all fluorescence-based imaging platforms
  • Conjugates to match any laser setting
  • Available in multicolor wavelengths for multiplexing applications
  • Available as stand-alone reagents or easy-to-use kits

Annexin V iFluor™ 555

Figure 4. Jurkat cells were treated with 1 µM camptothecin. The externalized PS residues, an early-stage indicator of apoptosis, was detected using Annexin V iFluor™ 555 conjugates (Cat No. 20072). Apoptotic cells and necrotic cells were were stained with Nuclear Green™ DCS1 (Cat No. 17550).

The table below shows the specifications of available annexin V iFluor™ conjugates to assist you in selecting the best annexin v conjugate for your fluorescence-based imaging or flow cytometry experiment.

 

Table 5. Available Annexin V iFluor™ Conjugates for Apoptosis Detection.

Annexin V Conjugate Ex (nm) Em (nm) Filter Set ε¹ Φ² Unit Size Cat No.
iFluor™ 350 344 344/448 DAPI 20000 0.95 100 tests (200 µL) 20070
iFluor™ 488 491 516 FITC 75000 0.9 100 tests (200 µL) 20071
iFluor™ 555 556 569 TRITC 100000 0.64 100 tests (200 µL) 20072
iFluor™ 594 587 603 Texas Red 180000 0.53 100 tests (200 µL) 20073
iFluor™ 633 638 652 Texas Red 250000 0.29 100 tests (200 µL) 20069
iFluor™ 647 654 669 Cy5 250000 0.29 100 tests (200 µL) 20074
iFluor™ 680 683 700 Cy5.5 220000 0.23 100 tests (200 µL) 20075
iFluor™ 700 690 713 Cy5.5 220000 0.23 100 tests (200 µL) 20077
iFluor™ 750 756 777 Cy7 275000 0.12 100 tests (200 µL) 20076
  1. Ext. Coeff. = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1).
  2. FQY = fluorescence quantum yield in aqueous buffer (pH 7.2).

 

Phalloidin iFluor™ Conjugates for Staining F-Actin

Used at nanomolar concentration, these iFluor™ phalloidin derivatives are convenient for many actin-related assays. Such assays include labeling, identifying, and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell-cultures, or cell-free experiments. Compared to traditional fluorescein isothiocynanate (FITC) and rhodamine conjugates, Phalloidin-iFluor™ conjugates offer F-actin stains that are superior in brightness and photostability. Phalloidin-iFluor™ conjugates are equivalent, and in most cases superior, to Alexa Flour® conjugates in performance.

Key features of Phalloidin iFluor™ Conjugates

  • Selectively targets and binds to F-actin, does not bind to monomeric G-actin
  • Compatible with formaldehyde-fixed samples and permeabilized tissue sections, cell cultures, cell-free experiments and paraffin-embedded samples that have been de-paraffinized
  • Negligible non-specific staining for high-contrast discrimination of actin
  • Available in multicolor wavelengths for multiplexing applications
  • Suitable for fluorescence-based imaging platforms

Phalloidin iFluor™ 594

Figure 5. F-actin of fixed and permeabilized HeLa cells visualized with Phalloidin iFluor™ 594 (Cat No. 23122). Endoplasmic reticulum (ER) and nuclei were stained, respectively, with Cell Navigator™ Live Cell ER Stainig Kit (Cat No. 22635) and DAPI (Cat No. 17507). ER was stained before fixation.

The table below lists specifications for Phalloidin-iFluor™ conjugates, their absorbance and emission maxima, extinction coefficient and fluorescence quantum yield.

 

Table 6. Phalloidin iFluor™ Conjugates For iFluor™ Bioconjugates

Conjugate Mol. Wt. Ex/ Em (nm) Filter Set Extinction Coefficient¹ Quantum Yield Unit Size
Phalloidin-iFluor™ 350 ∼1300 344/448 DAPI 20,000 0.95 300 Tests
Phalloidin-iFluor™ 405 ∼1400 402/425 DAPI 37,000 0.91 300 Tests
Phalloidin-iFluor™ 488 ∼1900 491/516 FITC 75,000 0.9 300 Tests
Phalloidin-iFluor™ 514 ∼1800 527/554 TRITC 80,000 0.83 300 Tests
Phalloidin-iFluor™ 532 ∼1800 543/563 TRITC 90,000 0.68 300 Tests
Phalloidin-iFluor™ 555 ∼1300 556/569 TRITC 100,000 0.64 300 Tests
Phalloidin-iFluor™ 594 ∼1600 587/603 Texas Red 180,000 0.53 300 Tests
Phalloidin-iFluor™ 633 1703.99 638/652 Texas Red 250,000 0.29 300 Tests
Phalloidin-iFluor™ 647 1632.87 654/669 Cy5 250,000 0.25 300 Tests
Phalloidin-iFluor™ 680 ∼2000 683/700 Cy5.5 220,000 0.23 300 Tests
Phalloidin-iFluor™ 700 ∼3000 690/713 Cy5.5 220,000 0.23 300 Tests
Phalloidin-iFluor™ 750 ∼3300 756/777 Cy7 275,000 0.12 300 Tests
Phalloidin-iFluor™ 790 ∼2800 786/811 Cy7 250,000 0.13 300 Tests
  1. Ext. Coeff. = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1).
  2. FQY = fluorescence quantum yield in aqueous buffer (pH 7.2).

 

Additional Resources:


 

Table 7. iFluor™ Dyes Spectral Properties For Additional Resources

iFluor™ Dye Abs. (nm) Em. (nm) Ext. Coeff.1 FQY2 CF at 260 nm3 CF at 280 nm4
iFluor™ 350 344 448 20,000 0.95 0.83 0.23
iFluor™ 405 402 425 37,000 0.91 0.48 0.77
iFluor™ 430 433 495 40,000 0.78 0.68 0.3
iFluor™ 450 451 502 40,000 0.82 0.45 0.27
iFluor™ 488 491 516 75,000 0.9 0.21 0.11
iFluor™ 514 527 554 80,000 0.83 0.25 0.11
iFluor™ 532 543 563 90,000 0.68 0.26 0.16
iFluor™ 546 541 557 100,000 0.67 0.25 0.15
iFluor™ 555 556 569 100,000 0.64 0.23 0.14
iFluor™ 560 559 571 130,000 0.57 0.048 0.069
iFluor™ 568 568 587 100,000 0.57 0.34 0.15
iFluor™ 594 587 603 180,000 0.53 0.05 0.04
iFluor™ 610 609 627 110,000 0.85 0.32 0.49
iFluor™ 633 638 652 250,000 0.29 0.062 0.044
iFluor™ 647 654 669 250,000 0.25 0.03 0.03
iFluor™ 660 660 677 250,000 0.26 0.07 0.08
iFluor™ 670 669 682 250,000 0.63 0.019 0.024
iFluor™ 680 683 700 220,000 0.23 0.097 0.094
iFluor™ 700 690 713 220,000 0.23 0.09 0.04
iFluor™ 710 712 736 190,000 0.14 0.12 0.07
iFluor™ 750 759 777 275,000 0.12 0.044 0.039
iFluor™ 790 786 811 250,000 0.13 0.1 0.09
iFluor™ 800 801 820 250,000 0.11 0.03 0.08
iFluor™ 810 811 822 250,000 0.05 0.09 0.15
iFluor™ 820 820 849 250,000 N/A 0.11 0.16
iFluor™ 840 836 876 200,000 0.02 0.2 0.09
iFluor™ 860 852 877 250,000 N/A 0.1 0.14
  1. Ext. Coeff. = molar extinction coefficient at their maximum absorption wavelength (Units = 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 (for oligo and nucleic acid labeling).
  4. CF at 280 nm is 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.

 

Table 8. iFluor™ Dyes Equivalents For Additional Resources

If you are using Try this iFluor™ Dye
Alexa Fluor® 350, AMCA, coumarin iFluor™ 350
Alexa Fluor® 488, Atto 488, HiLyte™ 488, Cy2, FITC iFluor™ 488
Alexa Fluor® 555, Atto 550, HiLyte™ 555, Cy3, TRITC iFluor™ 555
Alexa Fluor® 568, Atto 565, Rhodamine Red iFluor™ 568
Alexa Fluor® 594, Atto 594, Texas Red iFluor™ 594
Alexa Fluor® 647, Atto 647, HiLyte™ 647, Cy5 iFluor™ 647
Alexa Fluor® 680, Atto 680 Cy5.5, IRDye® 680 iFluor™ 680
Alexa Fluor® 750, HiLyte™ 750, Cy7 iFluor™ 750
  1. Alexa Fluor®is a registered trademark of ThermoFisher.
  2. HiLyte™ is a trademark of AnaSpec.

SUPPORT

outstanding technical support

PRODUCT

we offer a full product guarantee

DELIVERY

we offer free delivery to UK universities and non profit organisations