For alexa fluor 488, alexa fluor 532, alexa fluor 546, alexa fluor 555, alexa fluor 568, alexa fluor 594 and alexa fluor 647 dyes, qy measurements were made in pbs 50 mm potassium phosphate, 150 mm nacl, ph 7. This is a table of some characteristics of fluorochromes useful for flow cytometry or fluorescence microscopy. Alexa fluor fluoronanogold contains an extremely uniform 1. 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. It presents a list of imaginglabeling applications, biologicalmedical applications and industrial applications of alexa fluor 430 carboxylic acid succinimidyl ester. 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. Alexa fluor dyes spanning the visible and infrared. Alexa fluor 546 dye molecules can be attached to proteins.
Alexa fluor 532, alexa fluor 546, alexa fluor 555, alexa fluor 568, alexa fluor 594 and. An example of a fluorochrome with high quantum yield is alexa fluor 488 which has. The fate of this species is varied, depending upon the exact nature of the fluorophore and its surroundings, but the end result is deactivation loss of energy and return to the ground state. Fluoroquest fluorescence quantum yield determination kit optimized for bioconjugates catalog number. Fluorescence of alexa fluor dye tracks protein folding. In addition, unlike fluorescein, the fluorescence of the alexa fluor 488 dye is insensitive to ph between ph 4 and 10. Name max absorption max emission extinction coefficient quantum yield brightness index conditions.
Handbook of fluorescent dyes and probes wiley online books. Sulfonation makes alexa fluor dyes negatively charged and hydrophilic. F 1 signifies a 100% efficient fluorescence process. Alexa fluor 488 protein labeling kit thermo fisher scientific. The absorption maximum of alexa fluor 546 occurs near 556 nm, and the emission maximum is near 573 nm. Hilyte fluortm 488 501 527 alexa fluor514 518 540 evitag tm quantum dotscatskill green 525 540 alexa fluor430 434 541 pacific orange 403 551 alexa fluor532 532 554 hex 535 556 evitag tm quantum dotshops yellow 545 560 chromeo tm 546 545 561 cy3 548 561 488,514 alexa fluor555 555 565 hilyte fluortm 555 550 566 5tamra 541 568. F number of emitted photonsnumber of absorbed photons. Probes with high fluorescence quantum yield and high photostability allow detection of lowabundance biological structures with great sensitivity. The fate of this species is varied, depending upon the. The next generation fluorescent dyes for labeling proteins and. I molecules are photostable, prolongs the useful life of biosensors.
Alexa dyes have high quantum yield and are relatively photostable. Quantitative comparison of longwavelength alexa fluor dyes. The excitation and emission ranges of the alexa dyes cover the entire spectrum from ultraviolet to red, comprising wavelengths that match conventional light sources and including alexa 430 and alexa 532 dyes that, to our knowledge, represent the best dyes with good relative quantum yield. Alexa fluor 532 dye is phinsensitive over a wide molar range, helping ensure that you get stable signals in livecell imaging applications. The excitation and emission ranges of the alexa dyes cover the entire spectrum from ultraviolet to red, comprising wavelengths that match conventional light sources and including alexa 430 and alexa 532 dyes that, to our knowledge, represent the best dyes with good relative quantum yield in their excitation wavelength range. A 100% efficient fluorescence process would have a quantum yield of 1 the maximum quantum yield possible.
They are available in a wide spectral range from blue to deep red. The fluorescence quantum yield of alexa fluor 532 carboxylic acid, succinimidyl ester in 50 mm potassium phosphate, 150 mm nacl ph 7. Fluoroquest fluorescence quantum yield determination kit ordering information storage conditions instrument platform product number. Although it has remained difficult to separate the. Hilyte fluor 532 has extinction coefficient m 1 cm 1 171,000 and fluorescence quantum yield. 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 594 fluoronanogold our alexa fluor 594 fluoronanogoldconjugates offer an a second fluorophore, enabling the labeling of more than one target within a specimen with the brightness and photostability of the alexa fluor reagents. The alexa fluor 647 fluorophorelabeled oligonucleotides will not show visible fluorescence. This kit is only designed for a onepoint quick estimation of the fluorescence quantum yield of a biological conjugate. Pdf volume labeling with alexa fluor dyes and surface. Alexa fluor 532 is a fluorophore that exhibits strong. Fluorescence quantum yield is a measurement of emission efficiency and is calculated by the formula. For alexa fluor 488, alexa fluor 532, alexa fluor 546, alexa fluor 555, alexa fluor 568.
Alexa fluor 532 is a yellowfluorescent dye with excitation ideally suited for the frequencydoubled nd. Alexa fluor dyes are frequently used as cell and tissue labels in fluorescence. It presents a list of imaginglabeling applications, biologicalmedical applications and industrial applications of alexa fluor. Fluorescence of alexa fluor dye tracks protei n folding. Cf dyes rival or exceed the quality of other commercial dyes, such as alexa fluor dyes, due to the following novel features. Alexa fluor 430 carboxylic acid succinimidyl ester. Hilyte fluor 532 is suited for the frequencydoubled nd. Cf594, succinimidyl ester scj4600031 technical bulletin. Substance name solvent ph excitation max nm emission max nm qdot 705. Alexa fluor 532 dye is phinsensitive over a wide molar range, helping ensure that. Fluoroquest fluorescence quantum yield determination kit.
Cf594 also has excellent photostability, making it ideal for demanding applications such as confocal microscopy and single molecule imaging. Coeff e quantum yield qy brightness exqy100 commentsuse. The quantum yield is a readout of the efficiency of the fluorescence process. The alexa fluor series of dyes spans the nearuv, visible, and nearir spectrum, and are compatible with common excitation sources. They are conjugated postsynthesis to an aminomodified oligo as nhs esters. 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. Im currently using the alexa fluor 546 for the same purpose but it. Fluoronanogold combined fluorescent and gold nanoparticle. When conjugated to proteins, cf594 is significantly brighter than alexa fluor594 due to its high quantum yield and exceptional water solubility.
Alexa fluor 430 carboxylic acid succinimidyl ester handbook. Novel rhodamine chemistry rhodamine dyes are known for their excellent photostability and good fluorescence quantum yield. New rhodamine chemistry rhodamine dyes are known for their excellent photostability and good fluorescence quantum yield. Alexa fluor 647 dye is a bright, farredfluorescent dye with excitation ideally suited for the 594 nm or 633 nm laser lines. 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. Figure 2 comparison of relative quantum yield and total fluorescence of alexa fluor 555 gar and cy3 gar and of alexa fluor 647 and cy5 dyes conjugated to various proteins at different. Alexa fluor dyes are generally more stable, brighter, and less phsensitive than common dyes e. Pdf fluorescence of alexa fluor dye tracks protein folding. 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. 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. Quantitative comparison of longwavelength alexa fluor.
Laser 1 quantum brightness fluorochrome em nm filter lp. Within groups, roughly in order of excitation wavelength. Alexa fluor 488 dyelabeled proteins have absorption and fluorescence emission maxima of approximately 494 nm and 519 nm, respectively. A extinction coefficient of the acceptor m1 cm1 qy a quantum yield of the acceptor.
Alexa fluor 532 530 555 724 molecular probes alexa fluor 546 556 573 1079. Within groups, roughly in order of excitation wavelength families excepted. For alexa fluor 488, alexa fluor 532, alexa fluor 546, alexa fluor 555, alexa fluor 568, alexa fluor 594 and alexa fluor. Confocal microscopy list alexa fluor 532 with 543nm laser.
Alexa fluor dyes are frequently used as cell and tissue labels in fluorescence microscopy and cell biology. Human vision is insensitive to light beyond 650 nm, and therefore it is not possible to view the farredfluorescent dyes by looking. Amca conjugates, the fluorescence of alexa fluor 350 conjugates is better separated from that of. Fluorochromefluorescent proteinprobeindicator ex nm em nm mw oligomeric state mol. A guide to recording fluorescence quantum yields introduction. The alexa fluor 555 and the alexa fluor 647 dyes in protein conjugates exhibited signifi. Feb 04, 2014 however, there were outliers, such as alexa 633m. 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. For alexa fluor 488, alexa fluor 532, alexa fluor 546, alexa fluor 555, alexa fluor 568, alexa fluor 594 and alexa fluor 647 dyes, qy measurements were made in pbs. To overcome these problems, many nearir dyes, such as the nearir alexa fluor dyes, dylight dyes and irdyes, are engineered to have negatively charged sulfonate groups. Properties and structures a range of alexa dyes is available, with emission. Alexa dyes, a series of new fluorescent dyes that yield. The fluorescence quantum yield of alexa fluor 546 carboxylic acid, succinimidyl ester in 50 mm potassium phosphate, 150 mm nacl ph 7. Alexa fluor 532 spectrum alexa fluor 532 is a fluo.
When a fluorophore absorbs a photon of light, an energetically excited state is formed. The alexa, or alexa fluor, dyes were introduced by molecular probes, inc. For more accurate deamination of fluorescence quantum yield there are many factors that have a significant effect. Fluorochromes description cincinnati childrens hospital. Alexa fluor dyes sulfonation of coumarin rhodamine cyanine dyes. For stable signal generation in imaging and flow cytometry, alexa fluor. Laser 1 quantum brightness fluorochrome em nm filter lp e. 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. A lower yield may result from polymodifications andor strong secondary structures minimum guaranteed yields. Alexa fluor, pacific blue, and pacific orange antibody. Nano letters dependence of fluorescence intensity on the. An example of a fluorochrome with high quantum yield is alexa fluor. These postsynthesis manually coupled dyes are available in a wide spectral range from blue to deep red. Alexa fluor oligonucleotide amine labeling kits fluorophorelabeled oligonucleotides will show a visible fluorescence when illuminated with uv light, although this may be somewhat quenched by the presence of bromophenol blue.