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AAT Bioquest

mFluor™ Dyes and Kits

mfluor violet450
Live/Dead cell analysis using Annexin V-mFluor™ 450 conjugate
mFluor™ dyes are a series of superior fluorescent labeling dyes that span the UV to near-infrared (NIR) spectrum. Developed exclusively by AAT Bioquest, mFluor™ dyes exhibit excellent aqueous solubility and large Stokes Shifts, and their hydrophilic property minimizes the use of organic solvents. mFluor™ dyes have been extensively used to label antibodies proteins and other biomolecules for multicolor flow cytometry applications. The absorbance maxima of mFluor™ dyes are designed to be optimally excited by one of the major laser lines commonly equipped in flow cytometers, such as the 350 nm, 405 nm, 488 nm, 532 nm, 561-568 nm, or 633-647 nm laser lines. Compared to phycobiliproteins PE, APC, and their tandems, mFluor™ dyes are excellent alternatives for immunophenotyping, FACS, and other flow-based applications. mFluor™ dyes are available in a wide selection of products, including reactive dyes and antibody labeling kits, as well as mFluor™ streptavidin conjugates for signal amplification and annexin V-mFluor™ conjugates for apoptosis detection.



mFluor™ Reactive Dyes


mFluor™ reactive dyes can be covalently labeled to biomolecules without self-quenching, resulting in intensely bright fluorescent conjugates. mFluor™ reactive dye formats include amine-reactive succinimidyl esters (SE) and thiol-reactive maleimides for labeling antibodies and proteins. mFluor™ dyes are also available in acid form for labeling proteins, peptides, amine-modified oligonucleotides, and other amine-containing biomolecules by carbodiimide conjugation chemistry (EDAC).

mFluor Violet 450 conjugate
Flow cytometry analysis of HL-60 cells stained with 1 µg/mL mouse IgG control (Green) or with 1 µg/mL mouse anti-human HLA-ABC (W6/32 mAb) (Red) and then followed by goat anti-mouse IgG-mFluor™ Violet 450 conjugate. The fluorescence signal was monitored using an ACEA NovoCyte flow cytometer in the Pacific Blue channel.

Key features of mFluor™ reactive dyes

  • Simple, quick, and robust labeling of biomolecules with high conjugation yields
  • Superior brightness, photostability, and solubility
  • Strong fluorescence emission over a broad pH range with little pH sensitivity
  • Low spillover into most detectors
  • More choices and flexibility for multicolor panel design

The table below shows the range of available mFluor™ 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. For general antibody and protein labeling, use mFluor™ SE dyes which target primary amines (-NH2). For site-specific cysteine-directed conjugation, use mFluor™ maleimides.

 

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

Fluorophore
Laser (nm)
Ex/Em (nm)
Amine Reactive
Thiol Reactive
Alkyne Reactive
Carbonyl Reactive
Carbodiimide Reactive (EDAC)
mFluor™ UV375UV (350 nm)351/387mFluor™ UV375 SE    
mFluor™ UV420UV (350 nm)383/448mFluor™ UV420 SE    
mFluor™ UV455UV (350 nm)357/461mFluor™ UV455 SE    
mFluor™ UV460UV (350 nm)364/461mFluor™ UV460 SE    
mFluor™ UV520UV (350 nm)503/524mFluor™ UV520 SE    
mFluor™ UV540UV (350 nm)542/560mFluor™ UV540 SE    
mFluor™ UV610UV (350 nm)590/609mFluor™ UV610 SE    
mFluor™ Violet 450Violet (405 nm)406/445mFluor™ Violet 450 SEmFluor™ Violet 450 maleimidemFluor™ Violet 450 azide mFluor™ Violet 450 acid
mFluor™ Violet 480Violet (405 nm)404/475mFluor™ Violet 480 SE    
mFluor™ Violet 500Violet (405 nm)410/501mFluor™ Violet 500 SE    
mFluor™ Violet 505Violet (405 nm)393/504mFluor™ Violet 505 SE    
mFluor™ Violet 510Violet (405 nm)412/505mFluor™ Violet 510 SE   mFluor™ Violet 510 acid
mFluor™ Violet 540Violet (405 nm)402/535mFluor™ Violet 540 SE   mFluor™ Violet 540 acid
mFluor™ Violet 545Violet (405 nm)393/543mFluor™ Violet 545 SE    
mFluor™ Violet 550Violet (405 nm)527/550mFluor™ Violet 550 SE    
mFluor™ Violet 590Violet (405 nm)564/591mFluor™ Violet 590 SE    
mFluor™ Violet 610Violet (405 nm)421/613mFluor™ Violet 610 SE    
mFluor™ Blue 570Blue (488 nm)505/564mFluor™ Blue 570 SE   mFluor™ Blue 570 acid
mFluor™ Blue 580Blue (488 nm)485/580mFluor™ Blue 580 SE    
mFluor™ Blue 590Blue (488 nm)569/589mFluor™ Blue 590 SE    
mFluor™ Blue 620Blue (488 nm)589/616mFluor™ Blue 620 SE    
mFluor™ Blue 630Blue (488 nm)470/634mFluor™ Blue 630 SE    
mFluor™ Blue 660Blue (488 nm)481/663mFluor™ Blue 660 SE    
mFluor™ Green 620Green (532 nm)525/623mFluor™ Green 620 SE   mFluor™ Green 620 acid
mFluor™ Green 630Green (532 nm)537/657mFluor™ Green 630 SE    
mFluor™ Red 700Red (640 nm)680/695mFluor™ Red 700 SE   mFluor™ Red 700 acid
mFluor™ Red 780Red (640 nm)629/767mFluor™ Red 780 SEmFluor™ Red 780 maleimide mFluor™ Red 780 aminemFluor™ Red 780 acid

 


ReadiLink™ Rapid mFluor™ Antibody Labeling Kits from AAT Bioquest provide a convenient method for labeling microscale volumes of antibodies with our superior mFluor™ dyes. The unique chemistry of ReadiLink™ kits enables researchers to effortlessly label and recover 100% of their antibodies without a purification step. Since ReadiLink™ mFluor™ conjugates are covalently labeled, they are stable for long-term storage, and they are ideal for demanding applications including immunophenotyping, multiplex flow cytometry, FACS, and other flow-based applications. Also available are ReadiLink™ Rapid and xtra Rapid Antibody Labeling kits for conjugation of mFluor™ dyes, Alexa Fluor® dyes, other fluorescent dyes, biotin, BSA, and KLH.

ReadiLinkWorkflow

ReadiLink™ Rapid mFluor™ Antibody Labeling Kit workflow (figure drawn in BioRender).


Key features of ReadiLink™ mFluor™ Antibody Labeling Kits

  • 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
  • Conjugates ready to use in 1 hr, with as little as ~10 minutes of hands-on time
  • Avoid using any amine-containing buffers (e.g., Tris or glycine)
  • Stabilizing proteins (e.g., BSA or gelatin) must be removed before labeling; for antibody labeling kits compatible with BSA, use ReadiLink™ xtra Rapid Antibody Labeling Kits
Learn more

 

Table 2. Available ReadiLink™ Rapid mFluor™ Antibody Labeling Kits.

ReadiLink™ Rapid mFluor™ Labeling Kit
Laser Line (nm)
Ex/Em (nm)
Emission Filter (nm)
Imaging
Flow Cytometry
Cat No.
ReadiLink™ Rapid mFluor™ Violet 420405398/411425/20YesYes1105
ReadiLink™ Rapid mFluor™ Violet 450405406/445450/40YesYes1100
ReadiLink™ Rapid mFluor™ Violet 510405412/505525/50YesYes1110
ReadiLink™ Rapid mFluor™ Violet 540405394/537525/50YesYes1114
ReadiLink™ Rapid mFluor™ Blue 570488, 532, 561552/564575/26YesYes1120
ReadiLink™ Rapid mFluor™ Green 620488, 532, 561525/623610/20YesYes1123
ReadiLink™ Rapid mFluor™ Yellow 630488, 532, 561610/627610/20YesYes1126
ReadiLink™ Rapid mFluor™ Red 700633, 635, 640686/705730/45YesYes1130
ReadiLink™ Rapid mFluor™ Red 780633, 635, 640629/767780/60YesYes1131

 

mFluor™ Bioconjugates


We offer a large selection of mFluor™-labeled conjugates for a wide range of applications. Featured conjugates include Annexin V for apoptotic detection and streptavidin, which, when used together with biotin, can amplify weak signals in virtually any immunoassay.

Annexin V mFluor™ Conjugates for Apoptosis Detection


Annexin V mfluor™ Blue 570
The detection of binding activity of Annexin V-mFluor™ Blue 570 to phosphatidylserine in Jurkat cells. Jurkat cells were treated without (Green) or with 1 µM staurosporine (Red) at 37°C for 4 hours and labeled with Annexin V-mFluor™ Blue 570 conjugate for 30 minutes.
Annexin V conjugated to mFluor™ dyes provides a fast and reliable method for detecting externalized PS, a key characteristic in early-stage apoptosis. The superior brightness and photostability of Annexin V mFluor™ conjugates are useful for live-cell imaging, immunofluorescence, and flow cytometry. They can be combined with other dyes, such as nucleic acid stains, to accurately assess mixed populations of live, apoptotic, necrotic, and dead cells.

Key features of Annexin V mFluor™ Conjugates

  • mFluor™ 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

 

Table 3. Available Annexin V-mFluor™ conjugates for apoptosis detection.

Annexin V Conjugate
Ex (nm)
Em (nm)
Emission Filter (nm)
ε¹
Φ²
Unit Size
Cat No.
mFluor™ Violet 450406445450/40250000.92100 tests (200 µL)20080
mFluor™ Violet 510412505525/50300000.86100 tests (200 µL)20081
mFluor™ Violet 540394537525/50150000.64100 tests (200 µL)20082
mFluor™ Blue 570505564575/261200000.08100 tests (200 µL)20085
  1. ε = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1).
  2. Φ = fluorescence quantum yield in aqueous buffer (pH 7.2).

mFluor™ Streptavidin Conjugates for Signal Amplification


mfluor™ Yellow 630 streptavidin
Flow cytometry analysis of HL-60 cells stained with (Red) or without (Green) 1 µg/ml Anti-Human HLA-ABC-Biotin and then followed by mFluor™ Yellow 630-streptavidin conjugate.
Fluorescently labeled mFluor™ streptavidin conjugates have been widely used in fluorescence imaging to detect biotinylated antibodies, ligands, and DNA probes for in situ hybridization techniques, immunohistochemistry, multicolor flow cytometry, and other applications that require high sensitivity. mFluor™ streptavidin conjugates exhibit minimal self-quenching, producing exceptionally bright and photostable fluorescence signals outperforming Alexa Fluor® and other spectrally similar conjugates. In addition, the large Stokes shifts of mFluor™ dyes minimize cross-talk between the laser and emission filter and allow for cellular imaging with high signal-to-noise ratios. mFluor™ streptavidin conjugates are designed for optimal excitation by one of the major laser lines commonly equipped in flow cytometers, such as the 405 nm, 488 nm, 532 nm, 561 nm, or 633 nm laser lines.

Learn more

 

Table 4. Available mFluor™ streptavidin conjugates for flow cytometry.

 

Additional Resources


 

Table 5. Spectral properties of mFluor™ dyes.

mFluor™ Dye
Ex (nm)
Em (nm)
ε¹
Φ²
CF at 260 nm³
CF at 280 nm⁴
mFluor™ UV375351387300000.940.0990.138
mFluor™ UV460364461150000.860.350.134
mFluor™ Violet 450406445350000.920.3380.078
mFluor™ Violet 500410501250000.810.7690.365
mFluor™ Violet 510412505250000.860.4640.366
mFluor™ Violet 540402535N/D⁵N/D⁵N/D⁵N/D⁵
mFluor™ Blue 5705055641700000.08⁶0.2280.179
mFluor™ Violet 610421613920000.3N/D⁵0.66
mFluor™ Green 620525623500000.06⁶0.8950.569
mFluor™ Yellow 6306106271100000.01⁶0.2830.413
mFluor™ Red 7006806952950000.029⁶0.1350.127
mFluor™ Red 780629767900000.034⁶0.1010.116
  1. ε = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1).
  2. Φ = 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. N/D = not determined
  6. Fluorescence quantum yield may increase upon conjugation to antibodies.

Table 6. Fluorescent Dye Sets For Multicolor Applications

Cat#
Product Name
Unit Size
80051X4 Color™ B488 Dye Set 1 Set
80001X4 Color™ V405 Dye Set 1 Set

Table 7. mFluor™ Dye Equivalents

If you are using
Try this mFluor™ Dye
BD Horizon™ BUV395mFluor™ UV375
Pacific Blue®mFluor™ Violet 450
Hoechst 33342 or Marina Blue®mFluor™ UV460
Pacific Green™ or BD Horizon™ V500mFluor™ Violet 500
AmCyanmFluor™ Violet 510
Pacific Orange™ or Krome Orange™mFluor™ Violet 540
Phycoerythrin (PE)mFluor™ Blue 570
APC-Cy5.5®, APC-Alexa Fluor® 680, or APC-Alexa Fluor® 700 tandem dyesmFluor™ Red 700
APC-Cy7®, APC-Alexa Fluor® 750, or APC-H7 tandem dyesmFluor™ Red 780
  1. Alexa Fluor® is a registered trademark of ThermoFisher.
  2. Horizon™ is a trademark of BD Biosciences.

Table 8. Ordering Info for mFluor Products

Cat No.
Product Name
Unit Size
1616mFluor™ Violet 530 SE1 mg
1617mFluor™ Violet 530 maleimide1 mg
3111mFluor™ UV 375 Biotin Conjugate1 mg
3112mFluor™ UV 460 Biotin Conjugate1 mg
3113mFluor™ Violet 500 Biotin Conjugate1 mg
3114mFluor™ Violet 540 Biotin Conjugate1 mg
3115mFluor™ Red 780 Biotin Conjugate1 mg
3116mFluor™ Violet 450-PEG4-Biotin Conjugate1 mg
17011mFluor™ Violet 450-dUTP *1 mM in Tris Buffer (pH 7.5)*25 nmoles

 

Fluorescence Spectrum Viewer


Need assistance selecting the best fluorophore for your experiment, use our Fluorescence Spectrum Viewer:
  • View and compare fluorophores and fluorescent proteins for biological applications
  • Check spectral compatibility
  • Add multiple excitation and emission filters
  • Save spectra configuration as a PNG or hyperlink