Simplify your panel design with CF® Dyes
CF® Dye Advantages
- Available in over 30 colors, offering the greatest coverage across visible and near-IR wavelengths
- Widest selection of spectrally unique dyes to accommodate highly complex panels
- CF® Dye antibody conjugates have superior brightness and signal-to-noise over alternative fluorophores
Learn more about how CF® dyes offer advantages for enhanced multiplexing in spectral flow cytometry.
PRIMARY ANTIBODIES & ANTIBODY LABELING KITS
- Growing collection of more than 1000 monoclonal antibodies: View Flyer
- Choose from 13 bright CF® dyes, R-PE, APC, PerCP, HRP, AP, or biotin
- Isotype control antibodies available conjugated to the same dyes and labels
- Available BSA-free, ready to use for Mix-n-Stain™ labeling
- Affordable 100 uL sizes available
Primary Antibodies – sizes and formats:
|CF® Dye conjugates (17 colors)||0.1 mg/mL||100 uL or 500 uL|
|Biotin, HRP, or AP conjugates||0.1 mg/mL||100 uL or 500 uL|
|R-PE, APC, or Per-CP conjugates||0.1 mg/mL||250 uL|
|Purified, with BSA||0.2 mg/mL||100 uL or 500 uL|
|Purified, BSA-free (Mix-n-Stain™ Ready)||1 mg/mL||50 uL|
Streptavidin & Anti-Biotin Secondary Antibodies
Streptavidin and anti-biotin antibodies available in 21 bright CF® dyes, R-PE, APC, and PerCP
Antibody Labeling Kits
Labeling your own antibodies? Our Mix-n-Stain™ Antibody Labeling Kits dramatically simplify the process of preparing fluorescently-labeled antibodies, offering bright and stable conjugates in only 30 minutes. The resulting conjugates offer comparable performance to antibodies labeled with other commercially available labeling kits (Fig. 1.) Because the labeling is covalent, the conjugates are stable for long-term storage, and ideal for multi-color imaging and multiplex flow cytometry.
- Label with your choice of 29 CF® dyes in only 30 minutes
- Labeling Kits for FITC, Biotin, R-PE, APC, tandem dyes, and other labels also available
- Stable covalent labeling with no purification required
- Compatible with common antibody stabilizers
- Labeling kits designed for Nanobodies® (ie. camelid single variable or VHH domains) also available
Mix-n-Stain™ Tandem Dye Antibody Labeling Kits
Biotium’s novel APC and R-PE CF® tandem dyes are designed for optimal FRET between the donor and acceptor molecules for exceptional brightness and performance in flow cytometry.
- RPE-CF®583R (Ex/Em 496, 546, 586/609 nm) for the RPE-Texas Red® channel
- RPE-CF®647T (Ex/Em 496, 546, 555/665 nm) for the RPE-Cy®5 channel
- APC-CF®750T (Ex/Em 496, 546, 650/780 nm) for the APC-Cy®7 channel
- APC-CF®790T (Ex/Em 496, 546, 784/806 nm) for spectral cytometry or the APC-Cy®7 channel.
Learn more about our full selection of Mix-n-Stain™ Labeling Kits here.
LIVE-OR-DYE™ FIXABLE VIABILITY STAINING KITS
Live-or-Dye™ fixable dead cell stains are cell membrane-impermeant amine-reactive dyes. The dyes enter dead cells that have compromised membrane integrity and covalently label free amines on intracellular proteins (Fig. 3). Live-or-Dye™ labeling is extremely stable, allowing the cells to be fixed and permeabilized without loss of fluorescence or dye transfer between cells. The staining protocol has been optimized to maximize live/dead discrimination with minimal live cell staining, in order to prevent interference with immunostaining. Biotium currently offers a wide selection of 14 Live-or-Dye™ stains spanning the fluorescence spectrum for maximal flexibility in designing multi-color flow panels (Fig. 4).
The Best Fixable Dead Cell Stains for Flow Cytometry:
- Wide selection of dye colors for easy panel design
- Exceptionally bright dyes for excellent live/dead separation
- No loss of brightness after fixation
- For flow cytometry or fluorescence microscopy
- Dyes designed to fit gaps in spectral flow panels
- The most options for the 633 nm and 808 nm lasers
Spectral flow cytometry is a rapidly growing technology with significantly enhanced multiplexing capabilities over conventional flow cytometry. Where conventional flow cytometer instruments can detect panels with more than a dozen fluorophores, spectral flow cytometers can accommodate panels with upwards of 40 dyes. Biotium offers spectrally unique Live-or-Dye™ stains designed specifically to take advantage of enhanced multiplexing by spectral flow cytometry.
Live-or-Dye™ Stains for Spectral Flow
New: Live-or-Dye™ Sampler Kits
Live-or-Dye™ stains are available in two sampler kits, each containing five different dye colors, for excitation from all of the popular flow cytometry laser lines (see Live-or-Dye™ Sampler Kits table below). The Live-or-Dye™ Sampler Kit, Standard (32016) was designed for use on the most common flow cytometer laser and filter configurations, with dyes excitable by UV, Violet, Blue, Yellow-Green, and Red lasers. The Live-or-Dye™ Sampler Kit, Spectral (32017) was designed for use in spectral scanning flow cytometry. It contains dyes excitable by UV, Violet, Blue, and Red lasers, all of which have been validated on the Cytek® Aurora spectral cytometer, and chosen for their ability to fill the gaps in many spectral flow panels.
To learn more about Live-or-Dye™ Fixable Viability Stains, please visit the Live-or-Dye™ technology page.
Compatible lasers (nm)
Optimal detection channels
|32018, 32018-T||Live-or-Dye™ 330/410||355, 375||BUV395|
|32002, 32002-T||Live-or-Dye™ 350/448||355, 375||DAPI|
|32014,32014-T||Live-or-Dye™ 375/600||355, 375, 405||Spectral scan, BUV615, BV605||Developed for spectral cytometry|
|32003, 32003-T||Live-or-Dye™ 405/452||405||Pacific Blue®, BV421, BV450|
|32009, 32009-T||Live-or-Dye™ 405/545||405||AmCyan, BV510|
|32004, 32004-T||Live-or-Dye™ 488/515||488||FITC||Validated for microscopy|
|32012, 32012-T||Live-or-Dye™ 510/550||488, 532||Spectral scan||Developed for spectral cytometry|
|32005, 32005-T||Live-or-Dye™ 568/583||488, 532, 561||PE, PI||Validated for microscopy|
|32006, 32006-T||Live-or-Dye™ 594/614||488, 532, 561||PI, PE-CF®594, PE-Texas Red®||Validated for microscopy|
|32015, 32015-T||Live-or-Dye™ 615/740||633-640||Spectral scan, APC-Cy®7||Developed for spectral cytometry|
|32007, 32007-T||Live-or-Dye™ 640/662||633-640||APC||Validated for microscopy|
|32013, 32013-T||Live-or-Dye™ 665/685||633-640||Spectral scan, AF700||Developed for spectral cytometry|
|32008, 32008-T||Live-or-Dye™ 750/777||633-640, 785||APC-Cy®7||The brightest dye for the APC-Cy®7 channel|
|32011, 32011-T||Live-or-Dye™ 787/808||785, 808||APC-Cy®7, IR800|
Live-or-Dye™ Sampler Kits
Included dyes: Ex/Em (Cat No.)
|Live-or-Dye™ Fixable Viability Sampler Kit, Standard||32016||• 350/488 (32002A)|
• 405/545 (32009A)
• 488/515 (32004A)
• 568/583 (32005A)
• 640/662 (32007A)
|Designed for use on most standard flow cytometry laser and filter configurations|
|Live-or-Dye™ Fixable Viability Sampler Kit, Spectral||32017||• 350/448 (32002A)|
• 375/600 (32014A)
• 510/550 (32012A)
• 615/740 (32015A)
• 665/685 (32013A)
|Designed for use in spectral flow cytometry, to fill in gaps between common fluorophores|
Dead Cell Selective Counterstains
Cyanine-based cell membrane-impermeant nucleic acid stains such as Oxazole Blue (PO-PRO™-1), Oxazole Yellow (YO-PRO®-1), and Thiazole Red (TO-PRO®-3) are dead cell selective stains suitable for flow cytometry. Unlike Live-or-Dye™ Fixable Viability Stains, these cyanine-based dyes are not fixable, but are fluorogenic and highly specific for nucleic acids. This results in bright and sensitive staining that is localized to the nucleus in dead or dying cells. In addition, staining with these dyes can be done directly in cell culture medium without the need for washing. These dyes are generally known by their Thermo Fisher Scientific brand names and are available from blue to far-red. Biotium offers chemically equivalent forms of these dead cell selective stains at a high purity and lower cost.
|Oxazole Blue, 1 mM in DMSO||PO-PRO™-1||Blue (434/457 nm)||40091|
|Oxazole Blue Homodimer, 1 mM in DMSO||POPO™-1||Blue (433/457 nm)||40093|
|Oxazole Yellow, 1 mM in DMSO||YO-PRO®-1||Green (491/506 nm)||40089|
|Oxazole Yellow Homodimer, 1 mM in DMSO||YOYO®-1||Green (491/508 nm)||40090|
|TO Iodide, 1 mM in DMSO||TO-PRO®-1||Green (515/531 nm)||40088|
|Thiazole Orange Homodimer, 1 mM in DMSO||TOTO®-1||Green (514/531 nm)||40079|
|Oxazole Red, 1 mM in DMSO||YO-PRO®-3||Far-red (613/629 nm)||40105|
|Oxazole Red Homodimer, 1 mM in DMSO||YOYO®-3||Far-red (612/631 nm)||40106|
|Thiazole Red, 1 mM in DMSO||TO-PRO®-3||Far-red (642/657 nm)||40087|
|Thiazole Red Homodimer, 1 mM in DMSO||TOTO®-3||Far-red (642/661 nm)||40080|
Click the button on the right to view our our full selection of nuclear specific stains suitable for flow cytometry or microscopy.
VIAFLUOR® CELL PROLIFERATION
Cell Proliferation Monitoring by Flow Cytometry
Cell proliferation dyes diffuse passively into live cells and are used for long-term cell labeling. They are initially non-fluorescent esters of amine-reactive dyes, but are converted to fluorescent dyes by intracellular esterases. The dyes then covalently react with amine groups on intracellular proteins, forming fluorescent conjugates that are retained in the cell. Immediately after staining a single, bright fluorescent population will be detected by flow cytometry. Each cell division that occurs after labeling is revealed by the appearance of a successively dimmer fluorescent peak on a flow cytometry histogram. Cell proliferation dyes can be used to track cell divisions in vivo or in vitro. The staining can withstand fixation and permeabilization for subsequent immunostaining.
ViaFluor® SE Cell Proliferation Kits
CFSE (also known as CFDA-SE or carboxyfluorescein diacetate, succinimidyl ester) has been used for many years to monitor cell proliferation by flow cytometry. While several alternatives have been developed in more recent years, CFSE remains popular and widely used. Biotium continues to offer CFSE under the tradename ViaFluor® CFSE. However, CFSE has several drawbacks including leakage from the cell, cell toxicity, and bleed-through into the PE and PE-TexasRed® channels.
ViaFluor® 405 SE and ViaFluor® 488 SE cell proliferation dyes were developed at Biotium to provide superior cell staining, fixability, and low toxicity. ViaFluor® 405 is excited with the violet laser and detected in the Pacific Blue® channel, and gives great peaks with no toxicity. It behaves identically to the popular proliferation dye CellTrace™ Violet. ViaFluor® 488 and ViaFluor® CFSE are both detected in the FITC channel, but ViaFluor® 488 was developed as a less toxic, less leaky and more fixable alternative to the classic dye CFSE (Fig. 6). It also has less bleed-through into other channels such as PE and PE-Texas-Red®.
To learn more about ViaFluor® Cell Proliferation Kits, please visit the ViaFluor® technology page.
|ViaFluor® 405 Cell Proliferation Kit||30068||408/452||Pacific Blue®||• Track cell division by dye dilution using flow cytometry
• ViaFluor® 488 is a unique, improved green dye to replace CFSE
• ViaFluor® 405 replaces CellTrace™ Violet
|ViaFluor® 488 Cell Proliferation Kit||30086||493/532||FITC|
|ViaFluor® CFSE Cell Proliferation Kit||30050||495/519||FITC|
CELL CYCLE ANALYSIS
RedDot™1 Far-Red Nuclear Stain
RedDot™1 is a novel far-red nuclear stain developed at Biotium. RedDot™1
is a live cell stain similar to DRAQ®5 that can be used for cell cycle
distribution analysis (Fig. 7). In addition, RedDot™1 cell cycle analysis does
not require an RNase step unlike the classic nuclear stain Propidium Iodide
(PI). RedDot™1 can also be used as a far-red nuclear counterstain for live
cells in microscopy.
- Far-red cell membrane-permeant nuclear dye similar to Draq5™.
- Can be used for DNA content analysis by flow cytometry like Vybrant™ DyeCycle™ Ruby
- Highly thermostable and photostable, for convenient handling
NucSpot® 470 Nuclear Stain
NucSpot® 470 is a cell membrane-impermeant green fluorescent DNA stain for nuclear counterstaining of fixed cells, or selective staining of dead cells.
- Nuclear-specific green counterstain for fixed cells
- Selective detection of dead cells by flow cytometry in the FITC channel
- Perform cell cycle profiling by flow cytometry in fixed/permeabilized cells (Fig. 8)
- No wash required
NucSpot® Far-Red Nuclear Stain
NucSpot® Far-Red is an improved alternative to the popular flow cytometry dead cell dye 7-AAD. It has red-shifted fluorescence emission compared to 7-AAD, for less bleed-through fluorescence in the PE-Texas Red® channel.
- Improved alternative to 7-AAD with less bleed-through fluorescence in the PE-Texas Red® channel
- Selective detection of dead cells by flow cytometry in the PE-Cy®5 or APC channel
- Perform cell cycle profiling by flow cytometry in fixed/permeabilized cells (Fig. 9)
- Substitute for 7-AAD in any standard protocol
Dyes for Cell Cycle Analysis
|RedDot™1 Far-Red Nuclear Stain, 200X in Water||40060||Far-red|
|• For short-term live cell staining (≤4 hours)
• Analyze DNA content/cell cycle by flow cytometry
• Useful for cell number normalization for In Cell Western®
• Can be excited at wavelengths between 488 and 647 nm
• Detect in Cy®5 or APC channel
|NucSpot® 470 Nuclear Stain, 1000X in DMSO||40083||Green|
|• Nuclear-specific green counterstain for fixed cells
• Analyze DNA content/cell cycle by flow cytometry
• Selectively stains dead cells in live cultures
• Excellent match for blue LED excitation sources
|NucSpot® Far-Red, 1000X in DMSO||40085||Far-red|
|• Designed as improved replacement for 7-AAD
• For flow cytometry in the PE-Cy®5 or APC channel
• Less bleed into the PE-Texas Red® channel
|Propidium Iodide, 100 mg||40016||Red|
|• Widely used dead cell stain
• Can be excited by 488 nm laser line for detection in the PE channel by flow cytometry
• Useful for cell cycle analysis in fixed cells (with RNase treatment)
|Propidium Iodide, 1 mg/mL in Water||40017|
|Propidium Iodide Buffer, 50 ug/mL||40048|
|7-AAD, 1 mg||40037||Far-red|
|• Far-red dye for flow cytometry detection in the PE-Cy®5 channel
• Can be excited by the 488 nm or 532 nm laser line
• Useful for cell cycle analysis in fixed cells
|7-AAD, 1 mg/mL solution||40084|
Cy dye is a registered trademark of GE Healthcare. In Cell Western is a registered trademarks of LI-COR Inc.
NUCVIEW® CASPASE-3 SUBSTRATES
NucView® Caspase-3 Substrates for Live Cell Apoptosis Monitoring
NucView® caspase-3 substrates are novel fluorogenic substrates developed by Biotium to detect apoptosis in intact cells in real-time. Biotium has invented a new concept in the design of fluorogenic enzyme substrates. The enzyme substrate is attached to a fluorogenic DNA dye. Before cleavage, the dye is non-fluorescent and unable to bind to DNA. Upon enzymatic cleavage of the substrate, the dye is released and becomes capable of binding to DNA to emit fluorescence. In the case of NucView® caspase-3 substrates, the caspase-3/7 substrate peptide DEVD is attached to a DNA-binding dye. The substrate enters the cell cytoplasm where it is cleaved by caspase-3 in apoptotic cells to release the fluorogenic DNA dye, which stains the nucleus (Fig. 10). NucView® can be fixed with formaldehyde after staining, but it cannot be used to stain fixed cells or tissues.
- Simple, homogeneous assay for endpoint assay or real-time detection in live cells
- Non-toxic, allowing for multi-day experiments
- Detect caspase-3 activity and visualize apoptotic nuclear morphology
- For flow cytometry, fluorescence microscopy, or live cell imaging systems
- Available as stand-alone substrates or in kits with other apoptosis or necrosis probes
- Available with blue, green, or red fluorescence
- NucView® 488 validated in more than 100 cell types and 200 publications – See the list
To learn more about NucView® caspase-3 substrates, and kits please visit the NucView® technology page.
NucView® Caspase-3 Substrates and Kits
|NucView® 405 Caspase-3 Substrate, 1 mM in DMSO||10405||Blue fluorescence for flow cytometry in the Pacific Blue® channel or microscopy with the 405 nm laser|
|NucView® 405 Caspase-3 Substrate, 1 mM in PBS||10407||NucView® 405 substrate in PBS, for DMSO-sensitive cell types|
|NucView® 488 Caspase-3 Substrate, 1 mM in DMSO||10402||Green fluorescent substrate validated in more than 100 cell types and 200 publications|
|NucView® 488 Caspase-3 Substrate, 1 mM in PBS||10403||NucView® 488 substrate in PBS, for DMSO-sensitive cell types|
|NucView® 530 Caspase-3 Substrate, 1 mM in DMSO||10406||Orange fluorescence for microscopy in the Cy®3 channel or flow cytometry in the R-PE channel|
|NucView® 530 Caspase-3 Substrate, 1 mM in PBS||10408||NucView® 530 substrate in PBS, for DMSO-sensitive cell types|
|NucView® 488 and MitoView™ 633 Apotosis Detection Kit||30062||NucView® 488 and far-red fluorescent MitoView™ 633 for the Cy®5 channel|
|NucView® 488 and RedDot™ 2 Apoptosis & Necrosis Kit||30072||NucView® 488 and far-red dead cell DNA dye RedDot™ 2 for the Cy®5 channel|
|Dual Apoptosis Assay with NucView® 488 and CF®594 Annexin V||30067||NucView® 488 and red fluorescent Annexin V apoptosis probe|
|Dual Apoptosis Assay with NucView® 488 and CF®640R Annexin V||30073||NucView® 488 and far-red fluorescent Annexin V apoptosis probe|
CELL VIABILITY ASSAYS
Mitochondrial Membrane Potential
Loss of mitochondrial membrane potential is a hallmark for apoptosis. Biotium offers MitoView™ 633 dye for membrane potential-sensitive staining of mitochondria by microscopy or flow cytometry. For simultaneous detection of mitochondrial membrane potential and caspase-3 activity, see the NucView®488 and MitoView™633 Apoptosis Assay (Fig. 13).
We also offer MitoView™ 405, which changes localization upon mitochondrial depolarization, and MitoView™ Green, a membrane-potential independent mitochondrial dye that can be used to image mitochondria following mitochondrial depolarization, or after fixation.
In healthy cells, JC-1 dye aggregates in mitochondria as a function of membrane potential, resulting in red fluorescence with brightness proportional to the membrane potential. Conversely, in apoptotic and necrotic cells with diminished mitochondrial membrane potential, JC-1 exists in a green fluorescent monomeric form in the cytosol, allowing of cell viability to be assessed by measuring the ratio of red to green fluorescence by flow cytometry or fluorescence microplate reader. Rhodamine 123 is a green fluorescent mitochondrial membrane potential dye. Red fluorescent TMRM and TMRE are the preferred dyes for quantitative membrane potential measurements.
|MitoView™ 633||622/648 nm*||Cy®5, APC*||Yes||70055-T||50 ug|
|NucView® 488 & MitoView™ 633 Apoptosis Kit||500/530 nm (NucView®488)|
622/648 nm* (MitoView™ 633)
|MitoView™ 405||398/440 nm||DAPI||Partial†||70070-T||50 ug|
|MitoView™ Green||490/523 nm||FITC, GFP||No||70054-T||50 ug|
|JC-1 Mitochondrial Membrane Potential Detection Kit||510/527 nm (cytoplasm)|
585/590 nm (mitochondria)
|JC-1 Chloride Salt||70011||5 mg|
|JC-1 Iodide Salt||70014||5 mg|
|Rhodamine 123||505/534 nm||FITC||Yes||70010||50 mg|
|TMRE||548/573 nm||Cy®3||Yes||70016||25 mg|
|TMRE, 2 mM in DMSO||70016||0.5 mL|
|TMRM||548/573 nm||Cy®3||Yes||70017||25 mg|
*MitoView™ 633 also has red fluorescence in the Cy®3 channel and is not recommended for use with other red dyes.
Apoptosis/Necrosis Kits & Annexin V
Annexin V is a 36 kDa protein that has high affinity for phosphatidylserine (PS). During apoptosis, PS is translocated from the inner to the outer leaflet of the plasma membrane, where it can be stained by fluorescent conjugates of Annexin V. Annexin V conjugates are available with a wide selection of CF® dyes and other labels, including Near-IR CF® Dye Annexin V conjugates.
We offer many different staining kits with various combinations of dyes, including: NucView® caspase-3 substrate, our bright and photostable CF®488A Annexin V, a variety of dead cell nucleic acid stains, including propidium iodide (PI), 7-AAD, Ethidium Homodimer III (EthDIII), and RedDot™2. See the table below for more details on the various combination kits.
We offer dead cell nucleic acid stains separately as well, including far-red fluorescent RedDot™2 far-red dye and green fluorescent NucSpot® 470 stain.
Apoptosis and Necrosis Staining Kits
|CF®488A Annexin V and PI Apoptosis Kit||30061||• CF®488A stains apoptotic cells green for the FITC channel
• PI stains necrotic cells red for the Cy®3 or PE channels
• For microscopy or flow cytometry
|CF®488A Annexin V and 7-AAD Apoptosis Kit||30060||• CF®488A stains apoptotic cells green for the FITC channel
• 7-AAD stains necrotic cells red for the PE-Cy®5 or FL3 channels
• For flow cytometry
|Apoptosis & Necrosis Quantitation Kit Plus||30065||• CF®488A stains apoptotic cells green for the FITC channel
• EthD-III stains dead cells red for the Cy®3 or PE channels
• EthD-III has higher affinity and quantum yield than PI or 7-AAD
• For microscopy or flow cytometry
|Apoptotic, Necrotic & Healthy Cells Quantitation Kit Plus||30066||• CF®488A stains apoptotic cells green for the FITC channel
• EthD-III stains dead cells red for the Cy® channel
• Hoechst stains all cell nuclei blue for the DAPI channel
• For microscopy
|NucView®488 and RedDot™2 Apoptosis and Necrosis Kit||30072||• NucView®488 stains apoptotic nuclei green for the FITC channel
• RedDot™2 stains necrotic cells far-red for the Cy®5 or PE-Cy®5, APC, or FL3 channels
• For microscopy or flow cytometry
|Annexin V Conjugates||Multiple||• Annexin V with our bright and photostable CF® dyes and a selection of other labels|
|Annexin V Conjugates, Azide-Free||Multiple||• Annexin V with our bright and photostable CF® dyes and a selection of other labels
• Lyophilized and preservative-free for real-time cell imaging
|Annexin V Near-IR CF® Dye Conjugates||Multiple||• Annexin V conjugated to our superior Near-IR CF® dyes
• Lyophilized and preservative-free for in vivo use
|Annexin V (His Tag)||29088||• Recombinant Annexin V (His Tag) expressed from E.coli|
|Annexin Binding Buffer||99902||• Concentrated Annexin Binding Buffer for cell staining|
|NucSpot® 470 Nuclear Stain||40083||• Green fluorescent dead cell stain
• Also useful as a nuclear-specific counterstain for fixed cells
|RedDot™2 Far-Red Nuclear Stain||40061||• Far-red nuclear stain for dead cells or fixed cells
• Detect in the Cy®5 channel
|Ethidium homodimer III||40051||• Red fluorescent dead cell stain
• Higher affinity and quantum yield than PI
|Propidium Iodide||40017||• Widely used red fluorescent dead cell stain|
|7-AAD||40037||• Fluorescent dead cell stain for the PE-Cy®5/FL3 channel|
|NucSpot® Far-Red||40085||• Designed as an improved replacement for 7-AAD
• Less bleed into the PE-Texas Red® channel compared to 7-AAD