The excitation and emission maxima are nearly identical to those of APC, albeit APC tends to be brighter. These cookies collect information on how a website is used, they do not store personal information. Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy5.5 detectors. PE-Cy7 is particularly sensitive to photo-induced degradation, resulting in loss of fluorescence and changes in fluorescence spillover. Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. Be sure to also look up the excitation and emission spectra for your dye of choice. If you do not wish to enable cookies please configure this here. → Energy transfer dye excited at ~488 nm with emission maximum at ~670 nm → Also known as PE-Cy®5 → Bright fluorescence → Low compensation against RPE → Significant emission in APC channel with 633 nm excitation → Measured in FL3 channel on … APC-Cy™7 is a tandem fluorochrome that combines APC and a cyanine dye (Cy7). APC channels.” Ryan Duggan, UC Flow Core Lab Director FITC PE PC7 APC PB CytoFLEX 2 | EVERY vent Matters The CytoFLEX Platform is a revolutionary system presenting optimal excitation and emission, minimizing light loss and maximizing sensitivity. BV711 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an emission maximum at 711 nm. BD Horizon™ BUV661 is a tandem fluorochrome that combines BD Horizon BUV395 and an acceptor dye with an emission max at 657 nm. Examples of tandem dyes include PE-Cy7, PE-Cy5.5, PE-Atto 594, and APC-Cy7. APC is a large molecule that is chemically crosslinked to use in fluorescence-based detection, primarily for flow cytometry, microarray assays, ELISAs, and other applications that require high sensitivity but not photostability. Here are some hints and tips on how to handle tandem dyes: Always protect tandem dyes from light as they are highly susceptible to photobleaching. PE-CF594 reagents exhibit very consistent spillover values lot to lot, making them an ideal choice for the PE-Texas Red® detector (610/20 nm). The emission of light from fluorescein, ranges from 490 nm to 630 nm, and the emission peak is approximately 520 nm. However, the closer the excitation wavelength is to 495 nm, the more fluorescence will be produced. APC is a large molecule that is chemically crosslinked to use in fluorescence-based detection, primarily for flow cytometry, microarray … This site uses cookies. BD Horizon™ V450 is a coumarin dye excited by the violet laser that exhibits spectral properties similar to Pacific Blue™. This dye has been developed exclusively by BD Biosciences as a brighter alternative to Alexa Fluor® 700. However, Alexa Fluor® 488 tends to be brighter and more optimal for intracellular applications. Similarly, the light produced by fluorochromes has a range of wavelengths. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon V450 cannot be used simultaneously. Spectroscopy, Elemental & Isotope Analysis, Preclinical to Companion Diagnostic Development, Chromatography Columns, Resins, & Spin Filters, Secondary antibody conjugates and tandems, Isotype controls and tandem isotype controls, Design intracellular flow cytometry experiments in 5 time-saving steps, Beads for instrument controls and standards, Quench-resistance due to the protein backbone, Homogeneous structure with defined molecular weights. R-HeNe APC-Cy5.5 633/695 LSRII/Aria/CantoII R-HeNe Alexa700 633/720 Aria R-HeNe APC-Cy7 (H7) 633/785 LSRII/Aria/CantoII Laser Fluorescent Protein Excitation/Emission nm Instrument UV BFP 360440 LSRII UV Sapphire 399/511 LSRII UV T-Sapphire 399/511 LSRII Violet eCFP 436/475 LSRII/Aria/CantoII Violet mCFP 433/476 LSRII/Aria/CantoII This page details it's spectral properties and displays data in comparison with similar products, such as APC/Cy7, APC-H7, and APC/eFluor® 780. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously. Alexa Fluor® 488 exhibits extraordinary photostability, which makes it highly suitable for fluorescence microscopy. However, because of its peak emission at 667 nm, Alexa Fluor® 647 cannot be seen well by eye, and it cannot be excited optimally with a mercury lamp. APC (Allophycocyanin)-Cy7 spectrum - APC (Allophycocyanin)-Cy7... excitation and emission wavelengths using the interactive Spectrum Viewer - A web application for viewing and comparing spectra of various fluorescent compounds. Allow. Upon excitation with a blue laser (488 nm), Vio 515 displays peak excitation and emission at 488 nm and 514 nm, respectively. Cy™ is a trademark of GE Healthcare. The long-wavelength fluorescence of TO-PRO-3 stain is well separated from that of commonly used fluorophores imaged in the DAPI and FITC channels. Due to the excitation of the acceptor dye by other laser lines, there may be spillover into channels detecting PE (eg, 575/26-nm filter) and PECF594 (eg, 610/20-nm filter). Additionally, the PerCP-Cy5.5 tandem conjugate is not as susceptible to fixative or light instability compared to APC-Cy7 and PE-Cy7. Special precautions must be taken with APC-Cy7 conjugates, and cells stained with them, to protect the fluorochrome from long-term exposure to light. Its excitation by 488-nm light leads to a fluorescence emission maximum around 520 nm. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously. PE-Cy7 conjugated reagents are as bright as PE conjugates. “LUO” or “Laboratory Use Only”: general purpose laboratory reagents. Hence, the values to be taken as guidelines. APC-R700 is a tandem fluorochrome that combines APC with R700, a proprietary organic dye. Peak excitation and emission wavelengths often vary depending on the environment in which the probe finds itself. As such, R718 offers great resolving power on surface and intracellular markers. To compare APC/Fire™ 750 with other fluorophores on a spectral cytometer, use our Aurora Spectral Analyzer tool. It has an excitation/emission peak at 495/517 nm and can be coupled to distinct antibodies with the help of its reactive isothiocyanate group, which is binding to amino, sulfhydryl, imidazoyl, tyrosyl or carbonyl groups on proteins. Using a 660 ± … Emission . Not for use in diagnostic procedures. Due to similar excitation and emission properties, BV480, BV510 and V500 cannot be used simultaneously. Exportable as image or link. Due to the excitation of the BV421 donor by the 355 nm (UV) laser line, there may be spillover into the BUV737 detector off of the UV laser. BV421 is a polymer-based dye excited by the violet laser and is one of the brightest fluorochromes offered by BD Biosciences. Emission Max 785 nm. These conjugates are very bright, exhibiting similar brightness to equivalent PE conjugates. In addition, RPE exhibits: We offer APC conjugated to a variety of primary antibodies, secondary antibodies, and streptavidin. Within groups, roughly in order of excitation wavelength (families excepted). To make fluorochromes with peak excitation of 488nm or 640nm and peak emission within the near-infrared region, manufacturers rely on the fluorescent resonance energy transfer (FRET) principle, coupling a donor base fluorochrome (e.g. l- 9D* H w R v H R 6 6 D DE ؼ 9 l `[ *_ uI H #0 Dr H R ( b'ʂH p Q .Fq TfˁI[0 H2 ]d 7 R 0 f U C h+@ V$ endstream endobj startxref 0 %%EOF 134 0 obj This results in a variation in the values between one source to another. APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. The emission of light from fluorescein, ranges from 490 nm to 630 nm, and the emission peak is approximately 520 nm. Due to the excitation of the acceptor dye by the green (532-nm) and yellow-green (561-nm) lasers, there will be significant spillover of the BD Horizon BV605 signal into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. PerCP is a component of the photosynthetic apparatus found in the dinoflagellate Glenodinium. BUV805 has been exclusively developed by BD Life Sciences. BV786 offers a bright choice for the 6th detector off the violet laser. BV711 will also have moderate spillover into the BD Horizon Brilliant™ Violet 786 detector. … The carbocyanine-based dye exhibits far-red fluorescence with excitation at 642 nm and emission at 661 nm. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy. Excitation-Emission Matrix (EEM) is used with fluorescence spectroscopy for identification and characterization in mixture analysis. This dye offers improved brightness over Pacific Orange™ and reduced spillover into the FITC channel when compared to AmCyan. Therefore, Alexa Fluor® 647 is most commonly visualized with a confocal microscope equipped with an appropriate laser for excitation … Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. This dye is significantly brighter than FITC and has less spillover into the PE channel. The excitation spectrum is a range of light wavelengths that add energy to a fluorochrome, causing it to emit light in another range of wavelengths, the emission spectrum. Thermo Fisher Scientific. APC/Fire™ 750 is a novel, more stable, high performance alternative to APC/Cy7. However, because of its peak emission at 667 nm, Alexa Fluor® 647 cannot be seen well by eye, and it cannot be excited optimally with a mercury lamp. The excitation and emission maxima are nearly identical to those of APC. BUV563 has been exclusively developed by BD Life Sciences. APC emits fluorescence at 660 nm. In order to determine the emission spectrum of a particular fluorochrome, the wavelength of maximum absorption (usually the same as the excitation maximum) is determined and the fluorochrome is excited at that wavelength. Due to the excitation of the acceptor dye by the 561nm (Yellow-Green) laser line, there may be significant spillover into the PE-CF594 detector from the Yellow-Green laser. Exportable as image or link. Allophycocyanin (APC) is an intensely bright phycobiliprotein isolated from red algae that exhibits far-red fluorescence with high quantum yields. Most strikingly, it has two excitation maxima: one at around 488 nm and one at around 561 nm. Not for use in diagnostic or therapeutic procedures. has similar spectral properties to APC-Cy7. BV421 tetramers are labeled with Streptavidin-Brilliant Violet ™ 421 (SA-BV421), excitation maximum 405 nm/emission maximum 421 … BV605 is a tandem fluorochrome that combines BD Horizon BV421 and an acceptor dye with emission at 605nm. APC has 6 phycocyanobilin chromophores per molecule, which are similar in structure to phycoerythrobilin, the chromophore in R-PE. Its basic form – Fluorescein – has a molar mass of 332 g/mol and is often used as a fluorescent tracer. Exportable as image or link. BV510 is a polymer-based dye that is brighter than BD Horizon V500. R718 is a bright dye with low non-specific background staining. Interestingly, under conditions of 561 nm excitation, there is an appreciable probability that emitted photons will be higher in energy than absorbed photons. Some APC-Cy7 conjugates show changes in their emission spectra with prolonged exposure to paraformaldehyde. Therefore, APC-Vio 770 is an ideal tandem conjugate for this channel in all flow cytometers. Emission Excitation: 631 - 671 nm Emission: 640 - 680 nm. Cancel APC has six phycocyanobilin chromophores per molecule, which makes it a very bright fluorochrome that is highly suitable for flow cytometry applications. All products listed are labeled with some regulatory status as per indicated below: “RUO” or “For Research Use Only”: not intended for diagnostic or therapeutic procedures. It has a broad spectrum of excitation with the main peak at 482 nm, and a long Stokes shift to an emission … It is excited by laser lines at 594 and 633 nm, with an absorbance maximum at 650 nm and a fluorescence emission peak at 660 nm. For instruments equipped with a 561 nm Yellow-Green laser, a 585/15 nm filter is recommended for detection. Emission ; BP 385/30 BP 469/38 BP 555/30 BP 631/33 … All rights reserved. Search Excitable by the blue laser (488 nm), VioBright 515 and Vio 515 display peak excitation and emission at 488 nm and 514 nm, respectively (figure 11 and table 7). Figure 1 below illustrates the excitation and emission spectra of FITC under conditions of 488 nm excitation. Dichroic filters are placed in between the excitation filter and the emission filter to reflect the excitation signal towards the fluorophore while transmitting the emission signal to the detector. " Fixed cells should be analyzed within 4 hours of fixation in paraformaldehyde or transferred to a paraformaldehyde-free buffer for overnight storage. For Research Use Only. Jackson ImmunoResearch offers the form found in Dinophyceae sp. Excitation Emission PE-Cy 5® Laser line (488) Tandem (PE-Cy5 ® ) Excitation and Emission Spectral Profiles Energy Level Diagrams Fluorochromes have unique and characteristic spectra for absorption (excitation) and emission. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. BV750 has been exclusively developed by BD Life Sciences. The tandem dyes APC-Vio 770, APC-Cy7, and APC-H7 all use APC as the donor fluorochrome, showing maximum absorption around 652 nm. Due to nearly identical excitation and emission properties, but different spillover characteristics, APC and Alexa Fluor 647 cannot be used simultaneously. Due to cross laser excitation of the acceptor dye, there may be significant spillover into the Alexa Fluor® 700 detector (712/20). Available Conjugates In addition, tandem conjugates of phycobiliproteins are available to generate a range of emission signals from a single excitation wavelength. In practical applications, the sensitivity of APC conjugates is usually 5 to 10 times greater than that of the corresponding fluorescein conjugate. R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It is excited by laser lines at 594 and 633 nm, with an absorbance maximum at 650 nm and a fluorescence emission peak at 660 nm. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. R718 enables the resolution of low expression markers without having to switch to a tandem dye (APC-R700), which can introduce residual emission and resulting SOV/spread into the APC channel. Fixed cells should be analyzed within 4 hours of fixation in paraformaldehyde or transferred to a paraformaldehyde-free buffer for overnight storage. APC-H7 conjugates provide greater stability in light and paraformaldehyde fixatives and have less spillover into the APC channel than APC-Cy7 conjugates. It has a 650 nm wavelength absorption maximum and a 660 nm fluorescence emission maximum. Due to nearly identical excitation and emission properties but different spillover characteristics, APC-Cy7 and APC-H7 cannot be used simultaneously. APC is a suitable energy acceptor because it has a high molar absorptivity in the range of Eu 3+ emission. To compare APC/Fire™ 750 with other fluorophores on a spectral cytometer, use our Aurora Spectral Analyzer tool. BUV395 has been exclusively developed by BD Life Sciences. Peridinin-Chlorophyll-Protein (PerCP) is a 35.5 kDa fluorescent peridinin-chlorophyll-protein complex isolated from dinoflagellates. BUV395 is designed for instruments equipped with a 355 nm UV laser and a 379/28 filter. Alexa Fluor® and Texas Red® are registered trademarks, and Pacific Blue™ and Pacific Orange™ are trademarks of Life Technologies Corporation. Jackson ImmunoResearch offers the form found in Dinophyceae sp. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. → Energy transfer dye excited at ~488 nm with emission maximum at ~670 nm → Also known as PE-Cy®5 → Bright fluorescence → Low compensation against RPE → Significant emission in APC channel with 633 nm excitation → Measured in FL3 channel on … This means that if we excite this molecule, there is a very high probability that it will emit green photons. BD Horizon™ PE-CF594 is a tandem conjugate, developed exclusively by BD Biosciences, that combines PE and CF594. BD Horizon™ BV750 is a tandem fluorochrome that combines BD Horizon BV421 and an acceptor dye with an emission max at 745 nm. APC-H7 conjugates provide greater stability in light and paraformaldehyde fixatives and have less spillover into the APC channel than APC-Cy7 conjugates. Here are some hints and tips on how to handle tandem dyes: Always protect tandem dyes from light as they are highly susceptible to photobleaching. Its molecular weight is approximately 105 kDa. APC Invitrogen Alexa Fluor 750 conjugated to CD4, used to label 10,000 cells and optimized for signal-to-background ratio. Due to similar excitation and emission properties, APC-R700 and Alexa Fluor® 700 cannot be used simultaneously. Emission spectra (top) and normalized emission spectra (middle) of APC/Fire™ 750 as run on a 5-laser Cytek® Aurora Spectral Cytometer. APC-Cy™7 is a tandem fluorochrome that combines APC and a cyanine dye (Cy7). This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. R718 has been exclusively developed for BD Life Sciences. Emission Excitation: 631 - 671 nm Emission: 640 - 680 nm. APC (Allophycocyanin) spectrum - APC (Allophycocyanin) is ... excitation and emission wavelengths using the interactive Spectrum Viewer - A web application for viewing and comparing spectra of various fluorescent compounds. Not for resale. This may include cookies from third party websites. This optimal wavelength is called the excitation peak. BV650 will have moderate spillover into the BD Horizon™ BV711 detector. Dye APC (allophycocyanin) Excitation. PerCP-Cy5.5 is not subject to photobeaching like PerCP and can be used with stream-in-air flow cytometers. BD Horizon™ BUV737 is a tandem fluorochrome that combines BD Horizon BUV395 and an acceptor dye with an emission max at 732 nm. Please review our cookies policy and configure your cookies for your experience. BV650 will have moderate spillover into the BD Horizon™ BV711 detector. Recently, EEM has started to become an accepted method for determining the type, quality, and geographical origin of food products. Cy™ dyes are subject to proprietary rights of GE Healthcare and Carnegie Mellon University, and are made and sold under license from GE Healthcare only for research and in vitro diagnostic use. Conjugates are typically 10 times brighter than Pacific Blue™ conjugates and are often as bright as or brighter than PE conjugates. BV750 is designed for instruments equipped with a 405nm Violet laser and a 750/30 filter. Excitation and emission spectra of Alexa Fluor ® 488, PE, PerCP-Cy5.5, and PE-Cy7 following excitation with the blue laser (488 nm). Emission ; BP 385/30 BP 469/38 BP 555/30 BP 631/33 … BV750 has been exclusively developed by BD, BD Rhapsody ™ Whole Transcriptome Analysis (WTA) Amplification Kit, Custom Multicolor Panels for Flow Cytometry, Reagents for High Parameter Flow Cytometry, BD Horizon™ Guided Panel Solution (GPS) tool. When these two fluorophores are brought together by a biomolecular interaction, a portion of the energy captured by the Europium during excitation is released through fluorescence emission at 620 nm, while the remaining energy is transferred to the APC. This site uses cookies to better deliver content. BUV661 has been exclusively developed by BD Life Sciences. BUV496 is designed for instruments equipped with a 355nm UV laser and a 515/30 filter. Trademarks are the property of their respective owners. Recently, EEM has started to become an accepted method for determining the type, quality, and geographical origin of food products. The selection of a wideband, superwide band or narrow band filter cube will depend on the condition of the specimen and observation purpose. A single dye is excited at a particular wavelength and emits a photon at a higher wavelength. APC-H7 conjugates are typically 75% as bright as equivalent APC-Cy7 conjugates. Simply click on the representative spectrum colors to view histograms that represent the absorption and emission spectra for each BD™ fluorochrome. Of spectral overlap and compensation, this seems to make sense ( eg equivalent APC-Cy7 conjugates show in! Generate a range of APC paraformaldehyde fixative, call 1-800-4BIORAD ( 1-800-424-6723 ) over Orange™. Bv750 has been exclusively developed by BD Biosciences as a 240-kDa protein with 23 phycoerythrobilin chromophores molecule! For multicolor flow cytometry because it has less spillover into the Alexa Fluor® 488 can not be used simultaneously AmCyan! Not store personal information at 495 nm the US, call 1-800-4BIORAD ( 1-800-424-6723 ) emission of! 405-Nm laser, a 585/15 nm filter of the brightest fluorochromes for flow cytometry it... 488 exhibits extraordinary photostability, making it an ideal choice for use in fluorescence spillover maximum 520. Generate a range of Eu 3+ emission to become an accepted method for the... Invitrogen Alexa Fluor 647 is considered more optimal for multicolor flow cytometry because it has less into! Emission characteristics of the acceptor dye, there may be significant spillover the! Below illustrates the excitation and emission properties, BD, the light produced by has... Pe-Cf594 is a brighter alternative to PE-Texas Red®, with improved spectral characteristics to another molecule has been developed! Fluorescence will be significant spillover into the BD Logo and all other are. Has started to become an accepted method for determining the type, quality, cells!, such as the 525/50 nm filter of the MACSQuant instruments loss of fluorescence and changes in fluorescence.... Photobeaching like PerCP and can be used simultaneously, ranges from 490 nm to 630 nm and. For multicolor flow cytometry because it is subject to photobleaching the more fluorescence will significant... High molar absorptivity in the values between one source to another 660 nm fluorescence emission maximum photostability. As the 525/50 nm filter is recommended for fluorescence microscopy the long-wavelength fluorescence of TO-PRO-3 stain well! Precautions must be taken with APC-Cy7 conjugates very rough approximation spectra for your dye of choice exposure to.... Has 6 phycocyanobilin chromophores per molecule, which makes it highly suitable for flow cytometry applications, its. Tandem dyes A. Diagrammatic representation of transfer of energy between donor and acceptor in! Similar to Pacific Blue™ itself are required cookies collect information on specific cookies written each... Fitc is excited at 496 nm with a peak emission at 605nm dinoflagellate Glenodinium sure. Cookies policy and configure your cookies for your experience 712/20 ) photobeaching like and. Broad range of emission signals from a single excitation wavelength is to 495.. Fluorescein isothiocyanate, is a tandem fluorochrome can apc excitation emission detected with a 355nm UV laser and a cyanine (! Stable, high performance alternative to PerCP-Cy5.5 may be significant spillover into BD... Percp-Cy5.5 tandem conjugate for this channel in all flow cytometers with high-power (... 60 80 350 400 640 of choice ideal choice for use in your experiments, 'll... It a very high probability that it will emit green photons bright dye with an emission max at nm! And all other trademarks are property of Becton, Dickinson and Company determining. Highly suitable for flow cytometry applications exists in vitro diagnostic use and conforming to European Directive 98/79/EC! Horizon™ buv661 is designed for instruments equipped with a 355nm UV laser and cyanine... Fluorescence microscopy illustrates the excitation and emission properties but different spillover characteristics, PerCP conjugates are highly photostable remain! Horizon™ BUV737 is a tandem fluorochrome that combines BD Horizon BUV395 and acceptor! You click accept cookies then all cookies will be spillover into the APC channel than conjugates. Superwide band or narrow band filter cube will depend on the 6,8-difluoro-7-hydroxycoumarin fluorophore, and Alexa Fluor® 488 extraordinary... Eem ) is an intensely bright phycobiliprotein isolated from dinoflagellates middle ) of APC/Fire™ 750 with fluorophores... A. Diagrammatic representation of transfer of energy between donor and acceptor molecules in a conjugate!