The excitation and emission maxima are nearly identical to those of apc. Alexa fluor dyesacross the spectrum thermo fisher scientific fr. Design of multicolor flow cytometry panels incorporating. Welcome to the labeling chemistry selection tool featuring. Alexa fluor dyes are small molecules that react with free amino groups, primarily arginine and lysine huang et al. Alexa fluor dyes sulfonation of coumarin rhodamine cyanine. A wide spectrum of fluorescent dyes for your individual experiment delivered by. An entire panel of alexa fluor dyes emitting in the visible spectrum.
This dye offers improved brightness over pacific orange and reduced. However, information tends to be spread over many sites and is available in. The epub format uses ebook readers, which have several ease of reading. There is virtually no spectral overlap between these dyes and bleedthrough artifacts should be absent, even in specimens containing excessive levels alexa fluor. Alexa dyes, a series of new fluorescent dyes that yield. Also, the excited state lifetimes of the two dyes about 4 nsec each and the extinction coefficients of the two dyes. The alexa, or alexa fluor, dyes were introduced by molecular probes, inc.
Alexa fluor dyes are generally more stable, brighter, and less phsensitive than common dyes e. Our free online nucleic acids book contains information on all aspects of. Alexa fluor dyesacross the spectrum thermo fisher scientific kr. With the wide range of invitrogen alexa fluor dyes there is something for every filter and many options for multiplex detection. Bleedthrough makes it difficult to observe discrete fluorescence signals and. Hilyte fluor dyes are more photostable than the classic fluorescent dyes such as fitc, cy3 and cy5. Less nonspecific binding and less red spillover than alexa fluor 488.
Mechanisms of quenching of alexa fluorophores by natural. As members of dye families, such as alexa fluor 488, have been named numbered. Cf dyes are a series of highly watersoluble fluorescent dyes spanning the visible and nearinfrared nearir spectrum for labeling antibodies, proteins, nucleic acids. Alexa fluor dyes spanning the visible and infrared spectrum. Genecopoeia offers a full spectrum of andy fluor dyes for labeling antibodies, proteins, peptides, ligands, synthetic oligonucleotides and other biomolecules for use with immunochemistry, fluorescence in situ hybridization fish, cell tracing, receptor labeling, and cytochemistry applications as well as probing biological structure, function and interactions. Invitrogen alexa fluor dyes the leading and most trusted fluorescent dyes available today. Unfortunatly they didnt compare alexa fluor 647 with dylight 649 or as you mentioned alexa fluor 488 with dylight 488 but it shows the variability of dyes comming from one company based on there. The excitation and emission spectra of the alexa fluor series cover the visible spectrum.
In addition to superior performance youll get an experienced, problemsolving. Alexa fluor 488 491 515 1 mp a11001 ab label alexa fluor 546 553 569 1 mp a11003 ab label alexa fluor 594 594 610 1 mp a10239 ab label aminomethylcoumarin amc methylcoumarin 346. 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. Alexa fluor dyes have advantages compared to the cy dyes. The alexa fluor 660 dye is optimally excited with the 647 nm spectral line of the krypton ion laser and well excited by the 633 nm spectra line of the hene laser. A universal motif for fluorogenic azide probes across the. The alexa fluor dyes are highly water soluble, making them ideal for hybridization experiments. The alexa fluor dyes were chemically synthesized through sulfonation and additional chemical modifications made to the. The ctb conjugated with alexa fluor dyes can be obtained in the solid 500 ug. However, apc tends to be brighter while alexa fluor. Alexa fluor dyesacross the spectrum thermo fisher scientific. The current range of alexa fluor dyes available spans the visible spectrum with excitation wavelengths ranging from 346 to 784 nm. However, alexa fluor 488 tends to be brighter and more optimal for intracellular applications. Alexa fluor 647 antimouserathuman foxp3 antibody anti.
Oligonucleotides labeled with alexa fluor have many advantages. Review and cite alexa fluor protocol, troubleshooting and other. Performance relates to the molecular and steric similarities, which will vary for each new pair of dyes. The alexa fluor dyes have several properties that make them superior to other fluorescent dyes. Welcome to the labeling chemistry selection tool featuring invitrogen. The alexa fluor dyes are available from invitrogen. 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. Ive come across the clickit hpgalexa fluor azide and clickit aha alexa. Phalloidin binds factin with high selectivity while alexa fluor 350 provides blue fluorescence of unparalleled brightness and photostability. The wide range of alexa fluor dyes includes alexa fluor 350, alexa fluor 405, alexa fluor 430, alexa fluor 488, alexa fluor 514, alexa fluor 532, alexa fluor 546, alexa fluor 555, alexa fluor 568, alexa fluor 594, alexa fluor 610, alexa fluor 633, alexa fluor 635, alexa fluor 647, alexa fluor 660, alexa fluor 680, alexa fluor 700, alexa fluor 750, alexa fluor 790. The dye molecules were homogeneously distributed throughout the sio2 nanoparticles. Anaspec, egt groups proprietary ph insensitive hilyte fluor dyes are a series of excellent fluorescent labeling dyes with fluorescence emissions that span the full visible and near infrared spectrum.
Our fluorescent spectra graphingcalculator site presents dyes, filters, and. Bd horizon v500 is a novel organic dye excited by the violet laser. Less spillover fluorescence in the red channel than alexa fluor 488. Fluorescent dyes select the best label for your assay. Coeff e quantum yield qy brightness exqy100 commentsuse. Brightness brighter and more fluorescent than conventional dyes. Antia2b5 antibody, clone a2b5105 antibody, alexa fluor 647 conjugate. Thermo scientific dylight 488 and dylight 633 dyes. Their high quantum efficiencies make them suitable for single molecule studies where brightness is an important factor. Cy5, which emits in the far red range, provides a third colour for triple labelling. Absorption spectra of hilyte fluor dyes match the principal output wavelengths of common excitation sources such as 488 nm, 555 nm, 633 nm and 647 nm. The alexa fluor series of dyes spans the nearuv, visible, and nearir spectrum, and are compatible with common excitation sources.
Alexa fluor dyes from molecular probes thermo fisher. In this process, fluorescent alexa fluor dyes with different emission wavelengths were. The alexa fluor dyes bright, photostable fluorescent dyes from molecular probes for your research needs. Additionally, the water solubility of the dylight dyes allows a high dyetoprotein ratio without precipitation during conjugation. 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. Also make sure alexa 647 doesnt change its absorption spectrum in low ph i dont remember if it does. Fluorochromefluorescent proteinprobeindicator ex nm em nm mw oligomeric state mol. Ppt advantages of the alexa fluor dyes in molecular. Photostability 10 multiplexingthe four main colours emission wavelengths dapiuv fitc tritc far red alexa fluor 488 fluorescein fitc cy2 alexa fluor 555 rhodamine, tamra, tritc cy3 alexa fluor 647 cy5, apc alexa fluor 350 coumarin, amca alexa fluor. Biotium scientists discovered a new way to prepare novel rhodamine dyes of any. Over 30,000 published references and experienced technical support. Hilyte fluor dyes spanning the full visible spectrum.
Confocal microscopy spectral bleedthrough artifacts in. How do we remove free dye from a fluorescent labelled peptide. Nanowires replace newtons famous glass prism, university of cambridge. Perhaps the best spectral separation in visiblelight emitting alexa fluor dyes is the comparison between alexa fluor 488 and alexa fluor 633 depicted in figure 2c. Consult a spectrum viewer or a fluorophore reference chart to assess fluorophoremicroscope compatibility. Alexa fluor dyes are frequently used as cell and tissue labels in fluorescence microscopy and cell biology. Spans the nearuv, visible and nearir spectrum figure 1, table 1. The alexa fluor dyes were chemically synthesized through sulfonation and additional chemical modifications made to. A significant advantage of long wavelength dyes such as alexa fluor 647 is the low autofluorescence of biological specimens in this region of the spectrum. The spectral comparison i presented of alexa fluor 488 versus fluorescein was in a steady state fluorometer with relatively low power and nonsaturating conditions.
Alexa fluor 647 dye is a bright, farredfluorescent dye with excitation ideally suited for the 594 nm or 633 nm laser lines. Tandem dyes the power couples of flow cytometry biorad. Clickit edu imaging kit with alexa fluor 488, 594, and. Fluorescent bioorthogonal smart probes across the visible spectrum will. Alexa fluor dyes across the spectrum colour selection. Spectral imaging microscopy web sites and data mcnamara 2006. A simple method for alexa fluor dye labelling of dengue. 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.
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