To learn more about the different apoptosis pathways and the ways to detect and measure apoptosis in your cells, download this eBook from The Scientist and Biotium titled “Visualizing Apoptosis as it Happens”.
LIVE-OR-DYE™ FIXABLE VIABILITY STAINING KITS
Live-or-Dye™ Fixable Viability Staining Kits
Live-or-Dye™ fixable dead cell stains are cell membrane impermeable amine-reactive dyes. The dyes enter dead cells that have compromised membrane integrity and covalently label free amines on intracellular proteins (Fig. 1).
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 offers a selection of eight bright and photostable Live-or-Dye™ stains for maximal flexibility in multi-color analysis (Fig. 2). We also offer Live-or-Dye NucFix™ Red, a unique fixable nuclear-specific dead cell stain.
8 color options
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|
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. NucView® can be fixed with formaldehyde after staining, but it cannot be used to stain fixed cells or tissues.
- Simple, homogenous assay for endpoint assay or real-time detection in live cells
- Non-toxic, allowing for multi-day experiments
- Detect caspase-3 activity and visualize apopototic 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|
ANNEXIN V CONJUGATES
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 also offer Annexin V conjugates in an azide-free formulation, for long-term or in vivo studies.
|Biotin Annexin V||N/A||50 ug/mL||0.5 mL||Purchase 29013|
|CF®350 Annexin V||347/448 nm||50 ug/mL||0.5 mL||Purchase 29012|
|CF®405M Annexin V||408/452 nm||50 ug/mL||0.5 mL||Purchase 29009|
|CF®450 Annexin V||450/538 nm||50 ug/mL||0.5 mL||Purchase 29083|
|CF®488A Annexin V||490/515 nm||50 ug/mL||0.5 mL||Purchase 29005|
|CF®555 Annexin V||555/565 nm||50 ug/mL||0.5 mL||Purchase 29004|
|CF®568 Annexin V||562/583 nm||50 ug/mL||0.5 mL||Purchase 29010|
|CF®583R Annexin V||586/609 nm||50 ug/mL||0.5 mL||Purchase 29085|
|CF®594 Annexin V||593/614 nm||50 ug/mL||0.5 mL||Purchase 29011|
|CF®633 Annexin V||630/650 nm||50 ug/mL||0.5 mL||Purchase 29008|
|CF®640R Annexin V||642/662 nm||50 ug/mL||0.5 mL||Purchase 29014|
|CF®647 Annexin V||650/665 nm||50 ug/mL||0.5 mL||Purchase 29003|
|CF®660R Annexin V||663/682 nm||50 ug/mL||0.5 mL||Purchase 29069|
|R-PE Annexin V||496, 546, |
|5 uL/test||100 uL||Purchase 29045-100uL|
|500 uL||Purchase 29045-500uL|
|APC Annexin V||650/660 nm||5 uL/test||100 uL||Purchase 29057-100uL|
|500 uL||Purchase 29057-500uL|
|FITC Annexin V||496/516 nm||50 ug/mL||0.5 mL||Purchase 29001|
|5X Annexin V Binding Buffer||N/A||5X||15 mL||Purchase 99902|
FLUORESCENT VIABILITY STAINING KITS
Viability/Cytotoxicity Assay Kit
The Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells stains live cells with green fluorescence and dead cells with red fluorescence for detection by fluorescence microscopy or flow cytometry. Calcein-AM is a non-fluorescent cell-permeable ester that can passively enter cells. In viable cells, it is hydrolyzed by esterases in the cytoplasm to release the green fluorescent dye calcein. Calcein itself is membrane-impermeant and is only retained in viable cells with intact plasma membranes. Ethidium Homodimer III is a red fluorescent membrane-impermeant nucleic acid dye. It selectively stains dead cells with damaged plasma membranes (necrotic or late apoptotic cells).
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.
|CF®488A Annexin V and PI Apoptosis Kit||30061||• CF®488A Annexin V stains apoptotic cells green for the FITC channel
• PI stains necrotic cells red for the PE channel
|CF®488A Annexin V and 7-AAD Apoptosis Kit||30060||• CF®488A Annexin V stains apoptotic cells green for the FITC channel
• 7-AAD stains necrotic cells red for the PE-Cy®5 channel
|Apoptosis & Necrosis Quantitation Kit Plus||30065||• CF®488A Annexin V stains apoptotic cells green for the FITC channel
• EthDIII stains dead cells red for the PE channel
|Apoptotic, Necrotic & Healthy Cells Quantitation Kit Plus||30066||• CF®488A Annexin V stains apoptotic cells green for the FITC channel
• EthDIII stains dead cells red for the PE channel
• Hoechst stains all cell nuclei blue for the Pacific Blue channel
|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 APC channel
|Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells||30002||• Green dye Calcein AM detects intracellular esterase activity in live cells
• EthDIII stains dead cells red for the PE channel
MITOCHONDRIAL MEMBRANE POTENTIAL DYES
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.
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 visible red fluorescence in the Cy®3/rhodamine channel. It is not recommended for imaging with other visible red probes.
† Localizes to the cytoplasm after mitochondrial depolarization, but still retains fluorescence.
Cy Dye is a registered trademark of GE Healthcare.
VIAFLUOR® SE CELL PROLIFERATION KITS
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 (Figs. 3&4). 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 (Fig. 3). It behaves identially to the popular proliferation dye CellTrace™ Violet (Fig. 3). 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. 4). 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|