Quantitative comparison of longwavelength alexa fluor dyes to. The clickit tunel alexa fluor imaging assay contains all components needed to accurately and reliably detect apoptosis on adherent cells grown on coverslips or a 96well microplate, and includes dnase i to generate strand breaks as a positive control. Bacteriaconjugated alexa fluor dyes were resuspended into 1 x pbs to generate bacterial suspension at the concentration of 40. Volume labeling with alexa fluor dyes and surface functionalization of highly sensitive fluorescent silica sio2 nanoparticles. Quantitative comparison of longwavelength alexa fluor dyes. Quickly find the best reactive dye or label for your experiment. Mechanisms of quenching of alexa fluorophores by natural. Handbook of fluorescent dyes and probes is the most comprehensive volume available on the subject, covering all the available dyes and probes known to date in the literature for uses in various fields.
Compared to traditional dyes, alexa fluor dyes are brighter, more photostable, and more ph resistant between ph 4 and 10. Alexa fluor reactive dyes have good water solubility, so conjugations can be performed without organic solvents, and the conjugates are relatively resistant to precipitation during storage. Fluorescence excitation a and emission b of the dyes supplied with the fish tag dna kits. Are compatible with all common excitation sources table 2 and instruments. Handbook of fluorescent dyes and probes wiley online books. H igher sensitivity to elevate your image introducing an innovation in alexa fluor secondary antibodies offering higher signaltonoise ratio for your precious research samples providing up to 4x higher signaltonoise ratio, the unique, proprietary chemistry of new invitrogen alexa fluor plus secondary antibodies was designed to help you. Methods to covalently conjugate alexa fluor dyes to cellulose nanocrystals, at limiting amounts that retain the overall structure of the nanocrystals as model cellulose materials, were developed using two approaches. Jun 02, 2010 alexa fluor dyes from invitrogen are often used to label proteins, because they are more stable to photobleaching or conversions to the triplet states 2. This chapter lists the cas registry number, chemical structure, chemical abstract ca index name, other chemical names, merck index number, chemicaldye class, synthesis procedure, molecular formula, and molecular weight of the alexa.
Alexa 350, alexa 430, alexa 488, alexa 532, alexa 546, alexa 568, and alexa 594 dyes are a new series of fluorescent dyes with emissionexcitation spectra. The alexa fluor 660 dye is efficiently excited by the 633 nm and 647 nm spectral lines of the hene laser and the kryptonion laser, respectively. Quantitative comparison of longwavelength alexa fluor. Brightest dyes will be about as bright as pe while dim dyes will have brightness similar to bd horizon v500. It presents a list of imaginglabeling applications and biologicalmedical applications of alexa fluor 555 carboxylic acid succinimidyl ester. In addition to superior performance youll get an experienced, problemsolving technical support team, 50,000 published references, application and experimental tips, as well as protocols to help with experimental planning. Within groups, roughly in order of excitation wavelength families excepted. Alexa dyes, a series of new fluorescent dyes that yield.
Volume labeling with alexa fluor dyes and surface functionalization of highly sensitive fluorescent silica sio2 nanoparticles article pdf available in nanoscale 521. Also, various alexa fluor dyes are resistant to quenching at high degrees of substitution and thus are brighter than similar proteins labeled with other dyes. Alexa fluor dyes from invitrogen are often used to label proteins, because they are more stable to photobleaching or conversions to the triplet states 2. Properties and structures a range of alexa dyes is available, with emission wavelengths spanning the visible spectrum and extending into the infrared region of the spectrum. Alexa fluor 633 emits in the farred region which is well beyond the range of most sample autofluorescence alexa fluor 633 is bright. These postsynthesis manually coupled dyes are available in a wide spectral range from blue to deep red. Alexa fluor dyes can be conjugated directly to primary antibodies or to secondary antibodies to amplify. Pairs like alexa dyes 488555 and alexa dyes 488594 are often used. Dylight 550 562 576 4 alexa fluor 546, alexa fluor 555, cy3, tritc pe 496, 546 578 5 hfab rhodamine 530 580 3 texas red 596 615 2 apc 650 661 4 alexa fluor 647, cy5 alexa fluor 647 650 665 4 apc, cy5, dylight 650 cy5 649 670 3 dylight 650 654 673 4 alexa fluor 647, cy5 percp 490 675 2 starbright blue 700 470 700 5 dylight 680 692 712 4. Standard protein labelling procedure for fluidity onew.
Alexa fluor 555 carboxylic acid succinimidyl ester handbook. For example alexa fluor 488, phycoerythrin pe, peridinin chlorophyll protein percpcy5. Alexa 647 fret pairs with 40 and 30 bp, respectively. Alexa fluor 488 fitc 488 nm cy5 651 nm alexa fluor 647 wavelength nm 400 700 nm infrared fluorophores 700 nm uvvisible fluorophores popular applications. Molecular probes alexa fluor 647 protein labeling kit 1. Contents 4 your total resource for imaging 5 our commitment to quality and validation. Welcome to the labeling chemistry selection tool featuring invitrogen molecular probes reagents and labeling technologies. Volume labeling with alexa fluor dyes and surface functionalization of highly sensitive fluorescent silica sio2 nanoparticles article pdf available in nanoscale 521 september 20 with. The alexa fluor dyes, and other dyes that emit at longer wavelengths, towards the red end of the spectrum 600 nm have advantages over dyes that emit at shorter wavelengths in that there is typically less background fluorescence at longer these wavelengths. What are fluorecent probes and how do alexa fluor dyes work.
Relative fluorescence of cf543 and alexa fluor 546 af546 goat antimouse conjugates as a function of the number of dye molecules per protein degree of labeling. Aminereactive nhydroxysuccinimidyl esters of alexa fluor fluorescent dyes with principal absorption maxima at about 555 nm, 633 nm, 647 nm, 660 nm, 680. This is a table of some characteristics of fluorochromes useful for flow cytometry or fluorescence microscopy. Alexa fluor, pacific blue, and pacific orange antibody. Alexa fluor 633 632 647heliumneon laser 635nm alexa fluor is a series of dyes from molecular probes.
Developed by scientists using new breakthrough chemistries, the brightness, photostability, and color selection of cf dyes rival or exceed the quality of other. The benefits of alexa fluor dyes compared to traditional dyes, alexa fluor dyes are brighter, more photostable, and more ph resistant between ph 4 and 10. Relative photostability of cf633 and alexa fluor 647 af647 goat antimouse conjugates. It is important to know the filter sets available on your fluorescence microscope prior to choosing a fluorophore for labeling and detection table 2 and figure 1. These conjugates were compared with spectrally similar protein conjugates of the cy3, cy5, cy5. The alexa fluor family of fluorescent dyes is a series of dyes invented by molecular probes, now a part of thermo fisher scientific, and sold under the invitrogen brand name. Spans the nearuv, visible and nearir spectrum figure 1, table 1.
Alexa fluor dyes are typically used as cell and tissue labels in fluorescence microscopy and cell biology. One of the dyes that i want to talk about is the alexa fluor dyes. Organic dyes efluor organic dyes university of iowa. Molecular probes alexa fluor 647 antibody labeling kit 1. Green tea catechins quench the fluorescence of bacteria. Compounds used as dyes comparable to alexa fluor 350 dyes. Alexa dyes have high quantum yield and are relatively photostable. Pecf594 and alexa fluor 700 filters are available separately.
Green tea catechins quench the fluorescence of bacteriaconjugated alexa fluor dyes. Longrange singlemolecule forster resonance energy transfer. The dyes facilitate analysis of biological structures with enhanced sensitivity and selectivity. In the first, aldehyde groups are created on the cellulose surfaces by reaction with limiting amounts of sodium periodate, a reaction wellknown for oxidizing vicinal. Pdf green tea catechins quench the fluorescence of. Aminereactive nhydroxysuccinimidyl esters of alexa fluor fluorescent dyes with principal absorption maxima at about 555 nm, 633 nm, 647 nm, 660 nm, 680 nm, 700 nm, and 750 nm were conjugated to antibodies and other selected proteins. Alexa fluor dyes sulfonation of coumarin rhodamine cyanine. For protein labeling, the ability of alexa fluor dyes to retain their intense fluorescence even when the conjugates are heavily labeled suggests that the longwavelength alexa fluor dyes have advantages compared to the cy dyes and spectrally similar long. For details see alexa fluor dyes spanning the visible and infrared spectrumsection 1. Alexa 350, alexa 430, alexa 488, alexa 532, alexa 546, alexa 568, and alexa. Labeling chemistry selection tool welcome to the labeling chemistry selection tool featuring invitrogen molecular probes reagents and labeling technologies.
The alexa fluor 660 dye exhibits good spectral separation from orange fluorophores, including the alexa fluor 555 dye and phycoerythrin pe, making it a great choice for multicolor applications. Reactions with alexa fluor 546 are given as detailed examples, labeling on the order of 1% of the total glucopyranose rings of the cellulose. In addition, we prepare a variety of lysinefixable probes which have lysine residues incorporated into the dextran conjugate. The alexa, or alexa fluor, dyes were introduced by molecular probes, inc.
Alexa fluor dyes from molecular probes thermo fisher. Novel rhodamine chemistry rhodamine dyes are known for their excellent photostability and good fluorescence quantum yield. These modified dyes then were reacted with cellulose nanocrystals in acetonitrile at elevated temperature, again isolating material with the characteristic spectral bands of the alexa fluor dye. A large number of rhodamine dyes, including alexa fluor 488, alexa fluor 532, alexa fluor 568, atto488, atto532, and atto565, were found to form stable radical anions upon irradiation in the presence of thiol compounds such as. In experiments that involve cellscell debris, other chemicals e. Confocal microscopy fluorophores for confocal microscopy. Alexa fluorlabeled fluorescent cellulose nanocrystals for.
They are often used in cell and tissue labels when it comes to cell biology. And generally when using alexa fluor dyes, higher degrees of labeling can be achieved without intramolecular quenching. The alexa fluorlabeled antibodies can be used for multiple applications. The excitation and emission spectra of the alexa fluor series cover the visible spectrum and extend into the infrared. The inventive compounds have high fluorescence quantum yield and high photostability. Application filters mirrors alexa fluor 700 653446 cell surface markers 72030 705 lp bd horizon pecf594, petexas red 653447 618 605 lp gfp 653448 protein expression 51010 51010 yfp 653449 54520 535 lp optional mirror filter sets dyes application filters mirrors lp longpass filter class 1 laser product. Green dye that can be separated from cf488a by spectral unmixing. Tandem dyes are very useful for multicolor fluorescence studies, especially in combination with single dyes.
Alexa fluor dyes are available in a broad range of fluorescence excitation and emission wavelength maxima, ranging from the ultraviolet and deep blue to the nearinfrared regions. Their high quantum efficiencies make them suitable for single molecule studies where brightness is an important factor. Relative brightness is dependent on instrument configuration including lasers, filters, and laser power. Effects of egcg on the fluorescence of alexa fluor dyes. Which fluorophores are useful for flow cytometry biorad. The alexa fluor dyes are superior fluorophores with fluorescence emissions that. Standard protein labelling procedure for fluidity onew with fluorescent dye using alexa fluortm 488 nhs ester laboratory protocol from fluidic analytics ltd document reference. Link dye is a new generation of fluorescent probes that exhibit some advantages over other fluorescent dyes, such as being highly fluorescent over a broad ph range ph 4 to 10, good water solubility, and higher photostability, allowing more time for image capture. Molecular probes alexa fluor 647 antibody labeling kit 1 kit. In vivo imaging maximize the depth of tissue penetration using our comprehensive selection of infrared fluorescent in vivo imaging reagents. The alexa fluor family of fluorescent dyes is produced by molecular probes, a subsidiary of invitrogen. The majority of tandem dyes have been manufactured for the 488 nm and 640 nm lasers which are found in most cytometers. Pdf green tea catechins quench the fluorescence of bacteria.
Alexa fluor dyes antibody conjugates molecular probes alexa fluor 350. Alexa fluor 594 and alexa fluor 488 carboxylic acids invitrogen, carlsbad, ca, usa were dissolved in 1 x pbs at the concentration of 250 ngml. For details see alexa fluor dyes spanning the visible and. Alexa fluor dyes sulfonation of coumarin rhodamine cyanine dyes. Molecular probes offers numerous dextran conjugates in a variety of molecular weights and labeled with a wide array of substituents, including our superior alexa fluor dyes see tables 1 and 2. Pevio 770 pealexa fluor 750, pecy 7 apc alexa fluor 647, alexa fluor 700, apcalexa fluor 700, cy 5, draq5, efluor 660 apcvio 770 apcalexa fluor 750, apccy7, apcefluor 780, apch7, zombie nir table 2. Us8039648b2 compounds used as dyes comperable to alexa. Alexa fluor dyes are frequently used as cell and tissue labels in fluorescence microscopy and cell biology. Table of fluorochromes salk institute for biological studies. Tide fluor dyes for fluorescence labeling and fret. Cf dyes rival or exceed the quality of other commercial dyes, such as alexa fluor dyes, due to the following novel features. Invitrogen alexa fluor dyesthe leading and most trusted fluorescent dyes available today.
The alexa fluor dyes bright, photostable fluorescent dyes from molecular probes for your research needs. Alexa fluor dyes from molecular probes thermo fisher scientific. Covalently label proteins with fluorescent dyes which are optically compatible with the fluidity onew in a reliable and reproducible manner. Ram sabnis organizes the compounds alphabetically by the most commonly used chemical name. All efluor products are named for their emission wavelength and the organic dyes are fully compatible with proteinbased dyes, other organic dyes and efluor nanocrystal reagents. The chapter also includes references on the use and properties of alexa fluor 555 carboxylic acid succinimidyl ester.
Cf dyes are a series of highly watersoluble fluorescent dyes spanning the visible and nearinfrared nearir spectrum figures 1a and 1b for labeling antibodies, proteins, nucleic acids, and other biomolecules. Support material fluorescence spectra overview poster. Alphanumeric names of the individual dyes are associated with the specific excitation laser or arcdischarge lamp spectral lines for which the probes are intended. Apcefluor 780a tandem dye excited by the red 633 nm laser. Unlike other dyes, the fluorescence of alexa fluorconjugates is insensitive to ph between ph 4 and 10. Single dyes antibodies conjugated to fluorophores fluorophores fluorophore fluorescence color maximum excitation nm maximum emission nm relative brightness spectrally similar dyes dylight 405 400 420 3 alexa fluor 405, cascade blue alexa fluor405 401 421 3 pacific blue 410 455 1 dylight 488 493 518 4 alexa fluor 488, fitc. Alexa fluor 555 carboxylic acid succinimidyl ester. Alexa fluor 488 is ph independent between 410 fluorescein is significantly affected by ph. With the wide range of invitrogen alexa fluor dyes there is something for every filter and many options for multiplex detection. Pdf volume labeling with alexa fluor dyes and surface. Alexa fluor is a family of fluorescent dyes that can be purchased through the company fluoroprobes.