Part Number: 1225116 Price: $2,217 (Pricing is U.S. list. The filter combination utilizes wide-bandpass excitation and emission filters to provide maximum signal level. Autofluorescence emission intensity from starch grains and other components in a thin section of potato (Solanum tuberosum) tuber tissue is demonstrated in Figure 2(f). Metabion offers the Cy5.5 modification in its DNA and RNA portfolio. Available Conjugates A culture of bovine pulmonary artery endothelial cells was stained with phalloidin conjugated to Alexa Fluor 633, which targets the intracellular cytoskeletal F-actin network. Examine a culture of bovine pulmonary artery endothelial cells stained with MitoTracker Deep Red 633, which targets the intracellular mitochondrial network. Indodicarbocyanine, Cy5. Useful for most orange-emitting dyes such as TRITC. Invitrogen Cy5 dye is a bright, far-red-fluorescent dye with excitation ideally suited for the 633 nm or 647 nm laser lines. Endogenous autofluorescence in plant tissues arises from a variety of biomolecules, including chlorophyll, carotene, and xanthophyll. Fluorescence emission intensity from a culture of albino Swiss mouse embryo cells (3T3line) stained with the carbocyanine dye DiD, which targets lipophilic organelles, including the plasma membrane. The 2P-spectrum is sometimes included with the one-photon excitation (e.g. Because of its broad absorption range, PE-Cy5 is not recommended for use with dual-laser flow cytometers where excitation by both lasers is possible. This filter combination is optimized for use with popular fluorescent probes such as Cy5, Alexa Fluor 647, and allophycocyanin (APC), when the dyes are used alone or in multiple labeling techniques, and is also recommended for applications with other fluorophores excited by orange to red wavelengths. ♦ Excitation of CY 5.5 is about 675 (same as PerCP emission) and has a fluorescence emission maximum around 695nm APC-CY7 600-640nm 750-810Helium-Neon LASER (635nm) ♦ APC-CY7 is a tandem conjugate system, combining APC and a cyanine dye. Cy5 DBCO is an azide reactive probe used for imaging azide-tagged biomolecules via a copper-free “click reaction”. Observe the fluorescence emission intensity from a culture of canine kidney epithelial (Madin-Darby; MDCK line) cells stained with the carbocyanine dye DiD, which targets lipophilic organelles, including the plasma membrane. Autofluorescence emission intensity from starch grains and other components in a thin section of potato (Solanum tuberosum) tuber tissue. So, for example, PE-Cy5 will excite at the excitation wavelength for PE (565 nm) and emit at the emission wavelength for Cy5 (670 nm). The visible light absorption maximum of DiD is 644 nanometers and the emission maximum occurs at 665 nanometers. %PDF-1.6 %���� The Cy5 HYQ set employs a bandpass emission (barrier) filter (pass band region between 663 and 738 nanometers) instead of a longpass variety, in order to limit interference from fluorophores emitting outside the intended wavelength region, and to reduce autofluorescence. FAM Absorbance. Be sure to also look up the excitation and emission spectra for your dye of choice. Excitation is equivalent to absorption since upon absorption, the molecule reaches the excited state S n.The fluorescence excitation spectrum is obtained by fixing the emission wavelength and by running the excitation monochromator. �,� &��=ݠ6�#:�.P��X�!�� J�&s�jQH���(�DmJT�@��R4D��hq�4�RhkPT%�$*���P���N��8��4���l��}:I����_��}3'c�G�8}�M`�WqpR�E��a+�d��e��l�\����x���'�4�_�8�g7�|ٱ�^^&y2�$�s��x� n4�ϒˢ��ӫ��E����'L����n��9�b_6���u�$p���p��'�H�"���I�4Oq�l�����|��� a����W8�8�3�?�/��o���,\�5��fps���9,��;����?/*�T{��ZL�y�ȷ��~�/Z4p9�#��������l^log���(�%��"��0������"t|�o8<8��2~����l|�~;�M�c�����j�0'Er�j���\)/Q i����r�Q)�Ҿ4e�A�W��A�4_;�q�l�t)TҌø�`-��Q�������P�=�׋sI��.�Y�3���s.G`n]Uk)# z�UK}��r�ѼzN���H��/�O���s����h�__���� o{oN����1� ��|:��!�-f���͠� �Gл�OQ�I�'����Iv�P��?�1�Шd���B��$�. Fluorescence emission intensity from a culture of rat kangaroo (PtK2 line) epithelial kidney cells that were immunofluorescently labeled with primary anti-vimentin mouse monoclonal antibodies followed by goat anti-mouse F(ab')2 fragments conjugated to biotin. Cy™5 conjugates are excited maximally at 650 nm and fluoresce maximally at 670 nm. Ultraviolet, visible, and near-infrared transmission spectral profiles for components of the high-performance Nikon Cy5 HYQ filter combination are illustrated below in Figure 1. excitation and emission spectrum of Cy5. In the red absorption region, chlorophyll has an absorption band with a low extinction coefficient, but still produces a detectable level of fluorescence at emission wavelengths of 650 nanometers and above (deep red region). Mixture of Europium Cryptate Ubiquitin (SBB-TR0014), Cy5-Ubiquitin (SBB-TR0015), and wild-type Ubiquitin (SBB-UP0013) combined in a ratio optimized for TR-FRET based conjugation experiments where long polyubiquitin chains are formed. Note the bright fluorescence intensity exhibited by the intermediate filament network, which extends throughout the cytoplasm in these fibroblast cells. Fluorescence emission in a culture of rat kangaroo (PtK2 line) epithelial kidney cells that were immunofluorescently labeled with primary anti-vimentin mouse monoclonal antibodies followed by goat anti-mouse F(ab')2 fragments conjugated to biotin is demonstrated in Figure 2(c). Each individual component in this filter set is specifically designed to optimize detection of fluorescence from the fluorochrome Cy5 (along with spectrally similar probes), and the set also provides excellent performance for the majority of fluorophores excited by red wavelengths. The primary antibody, produced from purified nuclear fractions of HeLa cells, binds specifically to an antigen determinant that is present on all five histone proteins (H1, H2A, H2B, H3, and H4). for third color analysis because of its high emission intensity and broad spectrum. Atto dyes have strong fluorescent signals with most having molar absorptivity values >100,000 and low excitation/emission overlap, making Atto dyes ideal for multiplex techniques using visible and near-IR emission wavelengths. Emission filters are placed within the imaging path of a fluorescence microscope to pass only wavelengths within the emission range of the fluorophore. Phalloidins are a member of the phallotoxin group of bicyclic peptides isolated from the deadly Amanita phalloides mushroom. Cy5 dye. Nikon Instruments | Nikon Global | Nikon Small World. The visible light absorption maximum of Cy5 is 649 nanometers and the emission maximum occurs at 670 nanometers. The visible light absorption maximum of Alexa Fluor 633 is 621 nanometers and the emission maximum occurs at 639 nanometers. A single dye is excited at a particular wavelength and emits a photon at a higher wavelength. Home /; Technical Center /; Product Selection and Use /; Conjugate Selection /; Cyanine /; Cy5; Indodicarbocyanine, Cy™5. A longpass dichromatic mirror (660-nanometer cut-on wavelength) is provided in the Cy5 HYQ filter combination. This dye also has less cross-laser excitation off the 405-nm laser, resulting in less spillover into the violet channels compared to PerCP-Cy5.5. Figure 2(e) illustrates fluorescence emission in a culture of HeLa cells that were immunofluorescently labeled with primary anti-histone (pan) mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 647. Note the bright fluorescence intensity exhibited by the intermediate filament network, which extends throughout the cytoplasm in these epithelial cells. When excited by 488-nanometer light, the excited fluorochrome (PE) is able to transfer its fluorescent energy to the cyanine molecule, which then fluoresces at a longer wavelength in the red range at 670 nanometers. Spectral Properties . Note the bright fluorescence intensity exhibited by the fibrous vimentin intermediate filament network, which extends throughout the cytoplasm in these epithelial cells. For the starch grains illustrated above, note the presence of autofluorescence emission intensity in the red spectral region. ?��;�9�����o���~�I���|?�����l�o�pY�NOApy_C!҆K}ߖ�lHc���|��0^�!�Ukj���L%��n�X�U@dF!�[� �;X����j�C� ��$%ʰ܀�X�A��Y[T\5袢���FpO�a�7�4�҇��׭�LiC��ª���*"�@�DTX;V���Ayo�j�5O�X�[y���υ��ښ6q��pS��cm. 9 0 obj <> endobj 57 0 obj <>/Filter/FlateDecode/ID[<44A4923D141A8DC016CF3A2686ACE475><72D4CC2751DE4B20A141082DCE1B4DE3>]/Index[9 126]/Info 8 0 R/Length 182/Prev 749997/Root 10 0 R/Size 135/Type/XRef/W[1 3 1]>>stream Lifetime Data of Selected Fluorophores. Observe the fluorescence emission intensity from a culture of Indian Muntjac deerskin fibroblast cells that were immunofluorescently labeled with primary anti-cytokeratin (pan) mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 647. FAM Emission. The shopping cart is offline. Examine the fluorescence emission intensity from a culture of rat kangaroo kidney epithelial cells (PtK2 line) that were immunofluorescently labeled with primary anti-cytokeratin (pan) mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 647. (PE-Cy5 ® ) Excitation and Emission Spectral Profiles Energy Level Diagrams Fluorochromes have unique and characteristic spectra for absorption (excitation) and emission. J(λ) is the spectral overlap integral, given by: where φ D (λ) is the spectral shape of donor emission and ε A (λ) is the spectral shape of acceptor excitation (M −1 cm −1). GFP excitation - CY5 emission. Due to similar excitation and emission properties, BD Horizon BB700 and PerCP-Cy5.5 cannot be used simultaneously. Nikon offers a wide range of fluorescence filter cubes with high fluorescence acquisition efficiency to support imaging of a large variety of fluorophores and fluorescent proteins. Shipping: shipped on blue ice. A culture of male pig kidney epithelial cells (Sus scrofa; LLC-PK1 line) stained with MitoTracker Deep Red 633 is presented in Figure 2(b). Endogenous autofluorescence in plant tissues arises from a variety of biomolecules, including chlorophyll, carotene, and xanthophyll. Phycoerythrin-Cy5 (PE-Cy5, also written Cy5-PE) is a tandem conjugate where PE is coupled to the cyan dye, Cy5. Note the bright fluorescence intensity exhibited by the microtubule network in Figure 2(a). Chroma Catalog Sets: Cy3™/Cy5™ | | Included Filters: 51007x, 51007bs, 51007m | Related Fluorochromes: | This set utilizes the 546 nm line for Cy3™ excitation. An adherent culture of rat kangaroo kidney epithelial cells (PtK2 line) was immunofluorescently labeled with primary anti-cytokeratin (pan) mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Cy5. Cy5 HYQ - The Cy5 HYQ filter combination is the only Nikon set designed for high-energy red-wavelength excitation, and although optimized for use with the fluorophore Cy5, the wide yellow-orange to red excitation range is suitable for applications employing a variety of fluorochromes. They can be excited to about 98% of maximum with a krypton/argon laser (647 nm line) or to about 63% of maximum with a helium/neon laser (633 nm line). A significant advantage to using long wavelength dyes such as Cy5 dye or Invitrogen Alexa Fluor 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum. h޼YkS�J�+���hﻞ9sf��SRZ��g>�$< 1�c8���WZ��'t�Y{/�V�#im�g&b�0˙��,��J&�fV1a�� �O�i��p{�c�[��O(� A tandem dye consists of a Carbocyanine probes are often utilized in apoptosis experiments and as tracers for cellular migration. Cy5 dye. The filter combination utilizes wide passband excitation and emission filters to provide maximum signal level, as is characteristic of the Nikon HYQ filter combination group. "��l-�9D*��H�w R��v���H� R�6���6�D��DE�ؼ� ��9�� �l�`[��*_�uI�H #0�Dr���H��R��(�b'ʂH��p�Q�.Fq�TfˁI[0�H2���]d��7 ��R 0�f�� U��C�����h+@� �V$� endstream endobj startxref 0 %%EOF 134 0 obj <>stream Because they yield brighter and more stable fluorescence, cyanines can advantageously replace conventional dyes such as fluorescein and rhodamines. Fluorescence emission intensity exhibited by the tubular mitochondrial network extends throughout the cytoplasm of the central cell in the image. Caution: Excitation max and Emission max of the fluorescent molecules are subjected to the … Such sharp filter transitions are created by increasing the number of interference cavities in both the excitation and emission filters, enabling a closer spectral proximity between the two bandpass regions. The following data tables provide you with information on the fluorescent lifetimes, excitation and emission wavelengths of Selected fluorescent dyes, probes and labels that are frequently used for biological applications and in biomedical research. This far-red fluorescent probe is water-soluble, and its fluorescence is pH-insensitive from pH 4 to pH 10. The visible light absorption maximum of MitoTracker Deep Red 633 is 644 nanometers and the emission maximum occurs at 665 nanometers. The visible light absorption maximum of Alexa Fluor 647 is 649 nanometers and the emission maximum occurs at 666 nanometers (in the deep red region of the visible light spectrum). The Cy5 HYQ filter set is recommended when studying the following fluorophores: Alexa Fluor 633 and 647, APC, BODIPY 650/665, Cy5, Cy5.1.8, DiD (indodicarbocyanine), DiIC1(5), DiSC3(5), MitoTracker Deep Red 633, NeuroTrace 640/660, Nile Blue, SYTO (derivatives 60, 62, and 63), TO-PRO-3, and TOTO-3. The biotin secondary antibody was subsequently absorbed with a streptavidin conjugate of QDot 655. However, unlike the case with organic chromophores, fluorescence emission features a symmetrical profile in quantum dots (centered at 655 nanometers for this species). Carbocyanine probes are often utilized in apoptosis experiments and as tracers for cellular migration. Cy5 (Cyanine-5) spectrum - Cy5 (Cyanine-5) is a fluo... excitation and emission wavelengths using the interactive Spectrum Viewer - A web application for viewing and comparing spectra of various fluorescent compounds. The 75-nanometer emission window is combined with a wide 60-nanometer excitation passband (590 to 650 nanometers) to allow selective excitation and detection of specific popular fluorophores, which have absorption peaks in the orange to red spectral region. Anti-human golgin-97 antibodies recognize a 97-kiloDalton protein termed golgin-97, which is a member of the granin protein family and a peripheral membrane protein localized on the cytoplasmic face of the Golgi apparatus. h�b``�c``�a``��ia@�@���р,��=B�&*4+$1�u���]E��]�`�aV^�=xs��0&:�+�0�r�l� �#����`BA��;�a#���X(�9���y �$�nYޯsݥ+m@rw���j����_�3� �' endstream endobj 10 0 obj <> endobj 11 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text/ImageC]/Properties<>/Shading<>/XObject<>>>/Rotate 0/TrimBox[23 23 1587.724 2404.10205]/Type/Page>> endobj 12 0 obj <>stream Storage Conditions: store at -20 °C Short term exposure (up to 1 week cumulative) to ambient temperature possible. 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. BioTek Sample Files, 25-Mar-15, RFP excitation - CY5 emission, PN 1225117 The cells (3T3 line) were immunofluorescently labeled with primary anti-human golgin-97 mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Cy5. A version of the 89000 Quad set with a quad band excitation filter, dichroic and emission filter. For use with: Lionheart FX, Cytation 7, Cytation 5, Lionheart LX, Cytation 1. Within groups, roughly in order of excitation wavelength (families excepted). The Cy5 HYQ filter combination is designed with steep passband transitions, allowing the benefits of wide bandpass regions while maintaining sufficient signal separation to avoid an increased noise level relative to the detected emission. Peak excitation and emission wavelengths often vary depending on the environment in which the probe finds itself. Excitation and emission spectra of Alexa Fluor ® 488, PE, PerCP-Cy5.5, and PE-Cy7 following excitation with the blue laser (488 nm). Fluorescence emission intensity from a culture of male pig kidney epithelial cells (Sus scrofa; LLC-PK1 line) stained with MitoTracker Deep Red 633, which targets the intracellular mitochondrial network. h�bbd```b``� The bandpass emission filter functions to reduce or eliminate interference from emission in the infrared spectral region beyond approximately 740 nanometers. FAM Absorption and Emission Spectra . Compare the fluorescence emission intensity from the culture of rat kangaroo kidney cellular cytokeratin labeled with Cy5 to those listed above labeled with Alexa Fluor 647. The visible light absorption maximum of MitoTracker Deep Red 633 is 644 nanometers and the emission maximum occurs at 665 nanometers. ��8CDN0��R8�$�Z�b�s�LF�o��-S2��9�4�{��Q(�CҚ+˼`ZJμd�l�V{��i�%�����H�.5��:όQ�1�U�"ܯ�G[dIl#l�A`���yH/�yH/�yH/�y�eN�PW� �{�nPS�эě�n�S� �K�ɨg�*AաvI�ܡ~� a��G��-)[��Q}֢�B eG�R�U�;�>��H9 As with our PE conjugates, an average of one PE-Cy5 molecule is coupled per antibody or protein. The visible light absorption extinction coefficient of QDot 655 occurs over a wide range that increases with decreasing wavelength. A few examples: Alexa Fluors 350, 488, 568, 594, 610, 647, 660, 680, DAPI, DiI, various fluorescent proteins from the Drobizhev paper (see below), calcium indicators (incl. Canine kidney epithelial (Madin-Darby; MDCK line) cells were stained with phalloidin conjugated to Alexa Fluor 633, which targets the intracellular cytoskeletal F-actin network. Fluorescence emission intensity from a culture of HeLa cells that were immunofluorescently labeled with primary anti-histone (pan) mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 647. 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. The resulting wider and steeper filter passbands allow the set to provide more excitation energy and capture a higher level of fluorescence signal. Because the range of fluorescence wavelengths transmitted by the Cy5 HYQ combination extends into the near-infrared spectral region, which is either invisible or only faintly visible to the human eye, detectors with near-infrared sensitivity must be used to maximize the captured fluorophore emission. Local pricing may vary.) ♦ CY 7 has a fluorescence emission maximum around 767nm. Fura-2, Indo-1., Fluo3). A previous study demonstrated that a 1-μm solution of MCA-Pro-Leu (the product of enzymatic hydrolysis) showed excitation and emission at 328 nm and 393 nm, respectively, and was 130-fold more fluorescent than a comparable solution of MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Knight et al., 1992). Download the XML file for import into Gen5 Requires 465 nm LED cube p/n … Fluorescence emission intensity from a culture of Indian Muntjac deerskin fibroblast cells that were immunofluorescently labeled with primary anti-bovine alpha-tubulin mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Cy5. The Cy5 HYQ filter combination is designed to optimize detection of fluorescence from the fluorochrome Cy5, and the set also provides excellent performance for the majority of fluorophores excited by red wavelengths. We have measured the overlap integral for Cy3 and Cy5 terminally attached to dsDNA as J(λ) = 7.2 × 10 −13 M −1 cm 3. Broad UV Excitation 290-365 nm Emission Filters LP = Long pass filter DF = Band pass filter SP = Short pass filter 555 nm 555LP Molecular Imager FX Filters VersaDoc Filters 520 nm 520LP 660 nm 515 nm 530DF30 545 nm 622.5 nm 640DF35 657.5 nm 580 nm 605DF50 630 nm 670 nm 690DF50 500 nm 530DF60 560 nm 610 nm 610LP 660 nm Excitation Maxima Exportable as image or link. Get updates on our social media channels. 5' FAM-T10 oligonucleotide in PCR buffer . Cy5™ emission cannot be seen by … A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent chemical compound that can re-emit light upon light excitation. The two fluorescent probes (Alexa Fluor 647 and Cy5) have similar absorption and emission spectral profiles and produce comparable results with the Nikon Cy5 HYQ filter combination. May also be paired with individual emission filters from 89000 set for highest SNR image-splitting when acquiring multiple channels simultaneously. Emission Wavelength Range from 479 to 764 nm for Fluorescent Multiplex Detection. Cyanine 5 (Cy5) can be efficiently used in multiplex PCR in combinations. John D. Griffin, Nathan S. Claxton, Matthew J. Parry-Hill, Thomas J. Fellers, Kimberly M. Vogt, Ian D. Johnson, Shannon H. Neaves, Omar Alvarado, Lionel Parsons, Jr., Michael A. Sodders, Richard L. Ludlow, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310. Please call toll-free 1-866-SEMROCK (866-736-7625) or email us to place your order. Useful for simultaneous live viewing of multiple channels or image-splitting applications. Shelf Life: 12 months after date of delivery. Cy5. Invitrogen Cy5 dye is a bright, far-red-fluorescent dye with excitation ideally suited for the 633 nm or 647 nm laser lines. Electromagnetic energy from a laser set at the correct wavelength will provide the right amount of energy to an electron in the donor fluorescent dye molecule. Excitation Peak (nm) Emission Peak (nm) Cyanine, Cy2: 492: 510: Indocarbocyanine, Cy3: 550: 570: Indodicarbocyanine, Cy5: 650: 670 Anna Scordato and Stanley Schwartz - Bioscience Department, Nikon Instruments, Inc., 1300 Walt Whitman Road, Melville, New York, 11747. A culture of male pig kidney cells (Sus scrofa; LLC-PK1 line) was immunofluorescently labeled with primary anti-cytokeratin (pan) mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Cy5. DBCO moiety reacts with azides to form a stable triazole and does not require Cu-catalyst or elevated temperatures. For research use only! Molecular Formula: C 42 H 47 N 7 O 7 S 2. The fluorescence excitation spectrum characterizes the electron distribution of the molecule in the ground state. Presented in Figure 2(a) is the fluorescence emission profile from a culture of Indian Muntjac deerskin fibroblast cells that were immunofluorescently labeled with primary anti-bovine alpha-tubulin mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Cy5. All of the images in Figure 2 were captured in 12-bit grayscale mode, but subsequently pseudocolored by converting to the RGB color palette and setting the green and blue channels to zero (to produce a red image). A significant advantage to using long wavelength dyes such as Cy5 dye or Invitrogen Alexa Fluor 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum. Cy5, Cy5.5). Swiss albino mouse embryo cells (3T3 line) stained with the carbocyanine dye DiD, which targets lipophilic organelles, including the plasma membrane, are presented in Figure 2(d). Cy5.5 Cyanine 5.5 (Cy5.5) is a fluorescent dye that belongs to the Cyanine family of synthetic polymethine dyes. The images presented in Figure 2 demonstrate the performance of this filter combination with a variety of orange and red-absorbing fluorescence probes targeted at different intracellular locations. Usually, however, the two photon spectrum is indicated with -2P behind the fluorochrome's name. Cy5 is a fluorescent dye that belongs to the Cyanine family of synthetic polymethine dyes and it represents an excellent alternative to WellRED dye D4 and LIZ.