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CF®405S Dye

Outshines Alexa Fluor® 405

Brighter conjugates compared to Alexa Fluor® 405
Better match for common instruments
Validated for SIM super-resolution imaging
CF® Dye Spectrum:
CF®350 Dye

CF®350 is a blue fluorescent dye

300400500600700800900Wavelength (nm)CF350
Technical Summary

Abs/Em Maxima: 347/448 nm
Extinction coefficient: 18,000
Molecular weight: ~ 496
Excitation source: UV
Replaces: Alexa Fluor® 350, AMCA,
DyLight® 350

Features

Brighter and more photostable than AMCA
Direct replacement for Alexa Fluor® 350
Highly water soluble and pH-insensitive

CF®405S Dye

Improved brightness & photostability for the 405 nm laser line

300400500600700800900Wavelength (nm)CF405S
Technical Summary

Abs/Em Maxima: 404/431 nm
Extinction coefficient: 33,000
Molecular weight: ~ 1,169
Excitation laser line: 405 nm
Replaces: Alexa Fluor® 405, Cascade Blue®,
DyLight® 405

Features

Brighter than Alexa Fluor®  405
Validated for super-resolution imaging by SIM

CF®405M Dye

Improved brightness & photostability for the 405 nm laser line

300400500600700800900Wavelength (nm)CF405M
Technical Summary

Abs/Em Maxima: 408/452 nm
Extinction coefficient: 41,000
Molecular weight: ~ 503
Excitation laser line: 405 nm
Replaces: Pacific Blue®, BD Horizon™ V450

Features

More photostable than Pacific Blue®, with less spill-over in the green channel

An excellent choice for super-resolution imaging by SIM

CF®430 & CF®440 Dyes

Photostable 405 nm-excitable dyes with green fluorescence

300400500600700800900Wavelength (nm)CF430CF440
Technical Summary

CF®430
Abs/Em Maxima: 426/498 nm
Extinction coefficient: 40,000
Molecular weight: ~429
Excitation laser line: 405 nm
Replaces: Pacific Green®, BD Horizon™ V500,
Krome Orange™

CF®440
Abs/Em Maxima: 440/515 nm
Extinction coefficient: 40,000
Molecular weight: ~716
Excitation laser line: 405 nm
Replaces: Alexa Fluor® 430

Features

Photostable dyes suitable for microscopy
Perfect match for the CFP filter set
Highly water soluble and pH-insensitive
Suitable for flow cytometry in the AmCyan channel

CF®405L Dye

A 405 nm-excitable dye with green fluorescence emission

300400500600700800900Wavelength (nm)CF405L
Technical Summary

Abs/Em Maxima: 395/545 nm
Extinction coefficient: 24,000
Molecular weight: ~1573
Excitation laser line: 405 nm
Replaces: Pacific Orange®

Features

Expands potential detection channels for the 405 nm laser line

CF®450 Dye

A spectrally unique, 405 nm-excitable green dye

300400500600700800900Wavelength (nm)CF450
Technical Summary

Abs/Em Maxima: 450/538 nm
Extinction coefficient: 40,000
Molecular weight: ~689
Excitation laser line: 405 nm

Features

A spectrally unique, 405 nm-excitable green dye.

CF®488A Dye

A superior green fluorescent dye

300400500600700800900Wavelength (nm)CF488A
Technical Summary

Abs/Em Maxima: 490/515 nm
Extinction coefficient: 70,000
Molecular weight: ~914
Excitation laser line: 488 nm
Alternative for: Alexa Fluor® 488, DyLight® 488,
FITC, Cy®2

Features

Minimally charged, for less non-specific binding than Alexa Fluor® 488
Narrower emission spectrum for less bleed to red
Very photostable
Compatible with super-resolution imaging by TIRF
Highly water soluble and pH-insensitive

CF®503R Dye

A spectrally unique green dye ideal for spectral flow cytometry

300400500600700800900Wavelength (nm)CF503R
Technical Summary

Abs/Em maxima: 503/532 nm
Extinction coefficient: 90,000
Molecular weight: ~1100
Excitation laser line: 488 nm
Direct replacement for: ATTO 488

Features

A spectrally unique green dye ideal for spectral flow cytometry

CF®514 Dye

Alternative green fluorescent dye

300400500600700800900Wavelength (nm)CF514
Technical Summary

Abs/Em Maxima: 516/548 nm
Extinction coefficient: 105,000
Molecular weight: ~1216
Excitation laser line: 488 nm
Replaces: Alexa Fluor® 514

Features

Image using the same settings as FITC or CF®488A
Can be distinguished from CF®488A in the same specimen by spectral imaging and linear unmixing

CF®532 Dye

A bright green fluorescent dye for the 532 nm laser

300400500600700800900Wavelength (nm)CF532
Technical Summary

Abs/Em Maxima: 527/558 nm
Extinction coefficient: 96,000
Molecular weight: ~ 685
Excitation laser line: 532 nm
Direct replacement for: Alexa Fluor® 532, Atto 532

Features

Designed for the 532 nm laser
Brighter than Alexa Fluor® 532
Highly water-soluble and pH-insensitive

CF®543 Dye

An orange fluorescent dye ideal for the 543 nm laser

300400500600700800900Wavelength (nm)CF543
Technical Summary

Abs/Em Maxima: 541/560 nm
Extinction coefficient: 100,000
Molecular weight: ~ 870
Excitation laser line: 532 nm, 543 nm, or
546 nm
Direct replacement for: Alexa Fluor® 546,
TAMRA

Features

Optimized for the 543 nm laser
Yields the brightest conjugates among spectrally similar dyes
Highly water-soluble and pH-insensitive

CF®535ST Dye

A red fluorescent dye designed for STORM super-resolution imaging

300400500600700800900Wavelength (nm)CF535ST
Technical Summary

Abs/Em Maxima: 535/568 nm
Extinction coefficient: 95,000
Molecular weight: ~728
Excitation laser line: 532 nm

Features

Designed specifically for super-resolution imaging by STORM

CF®550R Dye

A spectrally unique orange/red dye ideal for spectral flow cytometry

300400500600700800900Wavelength (nm)CF550R
Technical Summary

Abs/Em maxima: 551/577 nm
Extinction coefficient: 100,000
Molecular weight: ~686
Excitation laser line: 532 nm or 568 nm

Features

A spectrally unique orange/red dye ideal for spectral flow cytometry

CF®555 Dye

A bright and photostable orange-red dye

300400500600700800900Wavelength (nm)CF555
Technical Summary

Abs/Em Maxima: 555/565 nm
Extinction coefficient: 150,000
Molecular weight: ~ 901
Excitation laser line: 532 nm or 568 nm
Alternative for: Alexa Fluor®
555, ATTO 550, Cy®3, DyLight® 549,
Rhodamine

Features

Brighter than Cy®3
Highly water-soluble
Validated in multicolor STORM super-resolution imaging

CF®568 Dye

Outshines Alexa Fluor®568

300400500600700800900Wavelength (nm)CF568
Technical Summary

Abs/Em Maxima: 562/584 nm
Extinction coefficient: 100,000
Molecular weight: ~ 714
Excitation laser line: 532 nm or 568 nm
Direct replacement for: Alexa Fluor® 568,
ATTO 565, Rhodamine Red

Features

Yields much brighter antibody conjugates than Alexa
Fluor® 568
Extremely photostable
Excellent choice for multi-color imaging with CF®488A and CF®640R
Compatible with TIRF and multicolor STORM super resolution imaging

CF®570

Bright red fluorescent dye

300400500600700800900Wavelength (nm)CF570
Technical Summary

Abs/Em maxima: 568/591 nm
Extinction coefficient: 150,000
Molecular weight: ~2998
Excitation laser line: 561 to 568 nm
Replaces: Alexa Fluor® 568, ATTO 565, DY-560, Rhodamine Red

Features

Produces antibody conjugates with brighter fluorescence compared to spectrally similar dyes

CF®583 & CF®583R Dyes

Bright red fluorescent dyes

300400500600700800900Wavelength (nm)CF583CF583R
Technical Summary

CF®583 Technical Summary
Abs/Em maxima: 583/606 nm
Extinction coefficient: 150,000
Molecular weight: ~3127
Excitation laser line: 561 to 568 nm
Alternative for: Cy®3.5

CF®583R Technical Summary
Abs/Em maxima: 586/609 nm
Extinction coefficient: 100,000
Molecular weight: ~773
Excitation laser line: 561 to 568 nm
Alternative for: Cy®3.5, Texas Red®

Features

Spectrally similar red fluorescent dyes with exceptional brightness
Serves as an alternative for Cy®3.5 and Texas Red®
Rhodamine-based CF®583R can be used as an excellent energy acceptor for FRET when paired with R-PE

CF®594 Dye

Truly the brightest deep red dye

300400500600700800900Wavelength (nm)CF594
Technical Summary

Abs/Em Maxima: 593/614 nm
Extinction coefficient: 115,000
Molecular weight: ~730
Excitation laser line: 561 to 568 nm
Replaces: Alexa Fluor® 594, DyLight® 594,
Texas Red®

Features

Yields the brightest antibody conjugates among spectrally
similar dyes.
Excellent choice for multicolor imaging with green dyes like CF®488A
Extremely photostable
Also see CF®594ST, a version of CF®594 engineered specifically for STORM microcopy

CF®594ST Dye

Deep red dye for STORM

300400500600700800900Wavelength (nm)CF594ST
Technical Summary

Abs/Em maxima: 593/620 nm
Extinction coefficient: 115,000
Molecular weight: ~868
Excitation laser line: 561 to 568 nm

Features

Designed specifically for super-resolution imaging by STORM

CF®597R Dye

Unique green-excited dye for STORM

4006008001,000Wavelength (nm)CF597R
Technical Summary

Abs/Em Maxima: 597/619 nm
Extinction coefficient: 115,000
Molecular weight: ~800
Excitation laser line: 561 to 568 nm
Replaces: Alexa Fluor® 594, DyLight® 594, ATTO 594

Features

Unique green-excited dye for STORM
Excellent solubility and photochemical switching properties
Validated for multicolor STORM with red-excited dyes

CF®620R Dye

A bright and photostable far-red dye

300400500600700800900Wavelength (nm)CF620R
Technical Summary

Abs/Em Maxima: 620/643 nm
Extinction coefficient: 115,000
Molecular weight: ~ 738
Excitation laser line: 633 nm or 635 nm
Replaces: LightCycler® Red 640

Features

Highly water-soluble
Highly fluorescent and extremely photostable
Can be used as an excellent energy acceptor for FRET when paired with R-PE

CF®633 Dye

The best dye for 633/635 laser lines

300400500600700800900Wavelength (nm)CF633
Technical Summary

Abs/Em Maxima: 630/650 nm
Extinction coefficient: 100,000
Molecular weight: ~820
Excitation laser line: 633 nm or 635 nm
Alternative for: Alexa Fluor® 633, Alexa Fluor®
647, Cy®5, DyLight® 633, DyLight® 649

Features

Yields the brightest antibody conjugates among spectrally similar dyes
Far more photostable than Alexa Fluor® 647
Compatible with TIRF, FIONA, and gSHRImP super-resolution imaging methods

CF®640R Dye

A highly photostable far-red dye

4006008001,000Wavelength (nm)CF640R
Technical Summary

Abs/Em Maxima: 642/662 nm
Extinction coefficient: 105,000
Molecular weight: ~ 832
Excitation laser line: 633 nm, 635 nm or
640 nm
Alternative for Alexa Fluor® 647, ATTO 647N,
Cy®5, DyLight® 649

Features

Best photostability among Cy®5-like dyes
Yields highly fluorescent protein conjugates
Compatible with TIRF and FLIMP super-resolution microscopy

CF®647 Dye

A highly fluorescent far-red dye

300400500600700800900Wavelength (nm)CF647
Technical Summary

Abs/Em Maxima: 650/665 nm
Extinction coefficient: 240,000
Molecular weight: ~ 1058
Excitation laser line: 633 nm, 635 nm or
640 nm
Alternative for: Cy®5, Alexa Fluor® 647,
DyLight® 649

Features

Brighter than Cy®5
Highly water soluble and pH insensitive
Validated in multi-color super-resolution imaging by STORM

CF®660C & CF®660R

Superior alternatives to Alexa Fluor® 660

300400500600700800900Wavelength (nm)CF660CCF660R
Technical Summary

CF®660C
Abs/Em Maxima: 667/685 nm
Extinction coefficient: 200,000
Molecular weight: ~ 3112
Excitation laser line: 633 nm, 635 nm or 640 nm
Replaces: Alexa Fluor® 660, APC

CF®660R
Abs/Em Maxima: 663/682 nm
Extinction coefficient: 100,000
Molecular weight: ~ 888
Excitation laser line: 633 nm, 635 nm or 640 nm
Replaces: Alexa Fluor® 660, APC

Features

CF®660C Features
Much brighter and more photostable than Alexa Fluor® 660
Compatible with multicolor super-resolution imaging by STORM

CF®660R Features
Brighter than Alexa Fluor® 660
Unrivaled photostability among spectrally similar dyes

CF®680 & CF®680R

Two outstanding 680 nm-excitable dyes

300400500600700800900Wavelength (nm)CF680CF680R
Technical Summary

CF®680
Abs/Em Maxima: 681/698 nm
Extinction coefficient: 210,000
Molecular weight: ~ 3241
Excitation laser line: 680 nm or 685 nm
Alternative for: Alexa Fluor® 680, Cy®5.5, IR®Dye 680

CF®680R
Abs/Em Maxima: 680/701 nm
Extinction coefficient: 140,000
Molecular weight: ~ 912
Excitation laser line: 680 nm or 685 nm
Alternative for: Alexa Fluor® 680, Cy®5.5, IR®Dye 680

Features

CF®680 Features
The brightest among spectrally similar dyes
Validated in multicolor STORM and 3D super-resolution
microscopy
Compatible with LI-COR® Odyssey®

CF®680R Features
Unrivaled photostability among spectrally similar dyes
Compatible with STED and single molecule spectroscopy super-resolution imaging (see pp. 20-21)
Molecular weight compatible with nucleic acid labeling
Compatible with LI-COR® Odyssey®

CF®700 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF700
Technical Summary

Abs/Em Maxima: 695/720 nm
Extinction coefficient: 240,000
Molecular weight: ~ 2315
Excitation laser line: 633, 635, 680 or 685 nm
Replaces: Alexa Fluor® 700, DyLight® 700

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging

CF®750 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF750
Technical Summary

Abs/Em Maxima: 755/777 nm
Extinction coefficient: 250,000
Molecular weight: ~ 3009
Excitation laser line: 633, 635, 680 or 685 nm
Alternative for: Alexa Fluor® 750, Cy®7, DyLight® 750

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging

CF®770 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF770
Technical Summary

Abs/Em Maxima: 770/797 nm
Extinction coefficient: 220,000
Molecular weight: ~ 3138
Excitation laser line: 785 nm
Replaces: DyLight™ 800, IRDye 800CW

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging

CF®790 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF790
Technical Summary

Abs/Em Maxima: 784/806 nm
Extinction coefficient: 210,000
Molecular weight: ~ 3267
Excitation laser line: 785 nm
Replaces: Alexa Fluor® 790

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging

CF®800 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF800
Technical Summary

Abs/Em Maxima: 797/816 nm
Extinction coefficient: 210,000
Molecular weight: ~3334
Excitation laser line: 785 nm
Spectrally similar to: Indocyanine Green

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging

CF®820 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF820
Technical Summary

Abs/EmMaxima: 822/835 nm
Extinction coefficient: 253,000
Molecular weight: ~2553
Excitation laser line: 785 nm

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging

CF®850 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF850
Technical Summary

Abs/EmMaxima: 852/870 nm
Molecular weight: ~2787
Excitation laser line: 808 nm

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Optimal for the 808 nm laser for flow cytometry

CF®870 Dye

Near-infrared CF® Dye

4006008001,000Wavelength (nm)CF870
Technical Summary

Abs/EmMaxima: 876/896 nm
Molecular weight: ~2773
Excitation laser line: 808 nm

Features

Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Optimal for the 808 nm laser for flow cytometry

CF® Dyes for Multi-Color Super-Resolution Microscopy

Technical Summary

Features

CF Dyes validated in SIM, STORM, STED, TIRF, and more
Superior brightness, photostability, and photochemical switching
Single label antibody conjugates for STORM
Growing list of publications

Figure 1. Normalized spectra of CF®405S goat anti-mouse conjugate in PBS.

CF®405S is a blue fluorescent dye with an absorption peak wavelength matching the 405 nm blue diode laser line (Figure 1). In addition, the emission peak wavelength of the dye is well-centered within the blue detection window of BD flow cytometers. As a result, flow cytometry analysis using CF®405S dye results in much brighter signal than that using Alexa Fluor® 405 (Figure 2). CF®405S also has been validated in super-resolution imaging by SIM. See CF®405S product lines below.

CF®405S Technical Summary

  • Abs/Em maxima: 404/431 nm
  • Extinction coefficient: 33,000
  • Molecular weight: ~1,169
  • Excitation laser line: 405 nm
  • Direct replacement for: Alexa Fluor® 405, Cascade Blue®, DyLight® 405
  • Fluorescence lifetime:
    τ (ns)/free acid in PBS pH 7.4, ε (ns) = 3.88 ± 0.05

Visit our FAQ for more fluorescence life time information on CF® dyes.

Figure 2. CF®405S yields brighter conjugates than Alexa Fluor® 405. Flow cytometry analysis in the Pacific Blue® channel of a BD LSRII of Jurkat cells stained with mouse anti-CD3 or isotype control, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor® 405 or CF®405S with varying numbers of dyes per antibody (degree of labeling, or DOL). Bars represent geometric mean fluorescence.
MCF-7 cells stained with rabbit anti-COXIV, followed by CF®405S goat anti-rabbit (mitochondria, blue) and mouse anti-pPol-II, followed by CF®647 goat anti-mouse (nuclei, red).

CF is a registered trademark of Biotium; Alexa Fluor, Cascade Blue, and DyLight are registered trademarks of Thermo Fisher Scientific.

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Faq

CF® Dyes

Most of our products are stable at room temperature for many days, so in all likelihood the product will still work just fine. To be on the safe side, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. One exception that we are aware of is GelGreen™, which is more sensitive to light exposure than most of our other fluorescent dyes. If GelGreen™ is exposed to ambient light for a prolonged period of time (days to weeks), its color will change from dark orange to brick red. If this occurs, the GelGreen will no longer work for gel staining.  


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See below for our recommended alternatives to Akoya tyramides. Unless noted as a direct replacement, these tyramide dyes are spectrally similar alternatives. They have not been validated as direct replacements for use with Akoya kits or imaging systems.

CF® Dye Alternatives for Akoya Tyramides

Akoya Tyramide Ex/Em (nm) Biotium Tyramide Ex/Em (nm) Biotium Cat. No. Notes CF® Dye Features
TSA Coumarin 402/443 CF®405S 404/431 92197 Much brighter & more
photostable alternative
CF®405S Features
Opal™ Polaris 480 450/500 CF®430 426/498 96053 Recommended
for abundant targets
CF®430 Features
Opal™520 494/525 CF®488A 490/515 92171 CF®488A Features
TSA Fluorescein 494/517 Fluorescein 492/514 96018 Direct replacement See CF®488A for a brighter &
much more photostable alternative
TSA TMR
TSA Cyanine 3
550/570 Cyanine 555 555/565 96020 Direct replacement
for Cyanine 3
See CF®555 & CF®568 for brighter
& more photostable alternatives.
CF®555 555/565 96021 Brighter & more
photostable alternatives
CF®555 Features
CF®568 562/583 92173 CF®568 Features
Opal™570 550/570 CF®550R 551/577 96077 CF®550R Features
CF®555 555/565
96021 CF®555 Features
TSA Plus Cyanine 3.5 581/596 CF®583R 586/609 96085 Brighter & more
photostable alternatives
CF®583R Features
CF®594 593/614 92174 CF®594 Features
Opal™620 588/616 CF®583R 586/609 96085 CF®583R Features
TSA Cyanine 5 648/667 CF®640R 642/662 92175 Brighter & more
photostable alternative
CF®640R Features
CF®647 650/665 96022 Brighter alternative CF®647 Features
CF®660R 663/682 92195 Less cross-talk with visible red dyes;
bright & extremely photostable
CF®660R Features
TSA Plus Cyanine 5.5
Opal™690
673/692
676/694
CF®680R 680/701 92196 Bright & extremely
photostable
CF®680R Features
Opal™ Polaris 780 750/770 CF®754 745/786 96090 Recommended
for abundant targets
Unique NIR dye for TSA
TSA Biotin N/A Biotin-XX N/A 92176 Direct replacement
TSA Plus DNP N/A DNP N/A 96019 Direct replacement
TSA Plus DIG N/A N/A N/A N/A N/A
Note: The CF® Dyes listed here are spectrally similar alternatives to Opal™ dyes, they have not been validated as direct replacements for use with Akoya's kits or imaging system. Opal is a trademark of Akoya Biosciences.  


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Bioscience kits The guaranteed shelf life from date of receipt for bioscience kits is listed on the product information sheet. Some kits have an expiration date printed on the kit box label, this is the guaranteed shelf life date calculated from the day that the product shipped from our facility. Kits often are functional for significantly longer than the guaranteed shelf life. If you have an older kit in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the kit still works for your application before processing a large number of samples or precious samples. Antibodies and other conjugates The guaranteed shelf life from date of receipt for antibodies and conjugates is listed on the product information sheet. Antibodies and other conjugates often are functional for significantly longer than the guaranteed shelf life. If you have an older conjugate in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. For lyophilized antibodies, we recommend reconstituting the antibody with glycerol and antimicrobial preservative like sodium azide for the longest shelf life (note that sodium azide is not compatible with HRP-conjugates). Chemicals, dyes, and gel stains Biotium guarantees the stability of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label. Fluorescent compounds should be protected from light for long term storage. If you have a Biotium compound that has been in storage for longer than one year that you wish to use, we recommend performing a small scale positive control experiment to confirm that the compound still works for your application before processing a large number of samples or precious samples. Expiration date based on date of manufacture (DOM) If your institution requires you to document expiration date based on date of manufacture for reagents, please contact techsupport@biotium.com for assistance. Chemical products with special stability considerations: Esters Ester compounds include the following: • Succinimidyl esters (SE, also known as NHS esters), such as our amine-reactive dyes • Acetoxymethyl esters (AM esters) such as our membrane-permeable ion indicator dyes • Diacetate-modified dyes, like ViaFluor™ 405, CFDA, and CFDA-SE cell viability/cell proliferation dyes Ester dyes are stable in solid form as long as they are protected from light and moisture. Esters are not stable in aqueous solution. Concentrated stock solutions should be prepared in anhydrous DMSO (see Biotium catalog no. 90082). Stock solutions in anhydrous DMSO can be stored desiccated at -20°C for one month or longer. Esters should be diluted in aqueous solution immediately before use. Succinimidyl esters (SE) should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound. A note on CF® Dye succinimidyl ester stability Succinimidyl esters (SE) are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Many commercially available fluorescent dyes used for life science research are heavily sulfonated dyes which makes them particularly hygroscopic, worsening the hydrolysis problem. In addition, for several commercially available SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while the SE group in all of Biotium’s CF® Dyes is prepared from an aliphatic carboxylic acid. This structural difference reduces the susceptibility of CF® Dye SE reactive groups to hydrolysis, resulting in relatively stable reactive dyes with consistently higher labeling efficiency compared to other SE derivatives of other fluorescent dyes. Maleimides, MTS and thiosulfate dyes Like the succinimidyl ester dyes, these dyes are also susceptible to hydrolysis, although generally to a much lower degree. Thus, for long term storage, anhydrous DMSO is recommended for making stock solutions. Other reactive dyes Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dye’s functional group. See the product information sheets for specific reactive dyes more information. Coelenterazines and D-luciferin Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also can be used as a solvent to minimize evaporation. If the solvent evaporates, the coelenterazine will still be present in the vial, so note the volume in the vial prior to storage so that you can adjust the solvent volume to correct for evaporation if needed. Prepare working solutions in aqueous buffers immediately before use. Coelenterazines are stable for up to five hours in aqueous solution. Aquaphile™ coelenterazines are water soluble formulations of coelenterazines. They are stable in solid form when stored as recommended. Aquaphile™ coelenterazines should be dissolved in aqueous solution immediately before use. They are stable for up to five hours in aqueous solution. Note that coelenterazines are predominantly yellow solids, but may contain dark red or brown flecks. This does not affect product stability or performance. If your coelenterazine is uniformly brown, then it is oxidized and needs to be replaced. D-luciferin is stable in solid form and as a concentrated stock solution when stored as recommended; it is not stable at dilute working concentrations in aqueous solution. Prepare concentrated D-luciferin stock solutions (typically 1-100 mg/mL) in water, and store in aliquots at -20°C or below for six months or longer. Prepare working solutions immediately before use.


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