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.
|Catalog No.||Viability Dye||Laser line||Emission filter||Abs/Em Max||Applications|
|32002, 32002-T||Live-or-Dye 350/448||355 nm||DAPI or Violet||347/448 nm||Flow Cytometry|
|32003, 32003-T||Live-or-Dye 405/452||405 nm||Pacific Blue®||408/452 nm||Flow Cytometry|
|32009, 32009-T||Live-or-Dye 405/545||405 nm||AmCyan||395/545 nm||Flow Cytometry,
|32004, 32004-T||Live-or-Dye 488/515||488 nm||FITC||490/515 nm||Flow Cytometry,
|32005, 32005-T||Live-or-Dye 568/583||488 or 561 nm||PE||562/583 nm||Flow Cytometry,
|32006, 32006-T||Live-or-Dye 594/614||488 or 561 nm||PE-Texas Red®||593/614 nm||Flow Cytometry,
|32007, 32007-T||Live-or-Dye 640/662||633 or 640 nm||APC||642/662 nm||Flow Cytometry|
|32008, 32008-T||Live-or-Dye 750/777||633 or 640 nm||APC-Cy®7||755/777 nm||Flow Cytometry,
|32010, 32010-T||Live-or-Dye NucFix Red||488 or 532 nm||PE-Texas Red®||520/610 nm||Flow Cytometry,
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.
|Product||Catalog No.||Ex/Em (nm)||Flow detection channel||Features|
|ViaFluor® 405 Cell Proliferation Kit||30068/30068-T||408/452||Pacific Blue®||
|ViaFluor® 488 Cell Proliferation Kit||30086/30086-T||493/532||FITC|
|ViaFluor® CFSE Cell Proliferation Kit||30050||495/519||FITC|
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® 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|
CELLULAR STAINS FOR FLOW
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.
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.
|Product||Abs/Em||Detection channel||Potential-dependent?||Catalog no.||Size|
|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|
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.
|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 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 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|
PRIMARY & SECONDARY ANTIBODY CONJUGATES
- Wide selection of host and target species and anti-tag antibodies
- Your choice of more than 20 CF® dye colors, R-PE, APC, HRP, AP or biotin
- Highly cross-adsorbed, F(ab’)2 fragments, and isotype-specific options
- Affordable 50 uL sizes available
To learn more about our primary and secondary antibodies, and kits please visit the Antibodies technology page.
Primary Antibodies – your choice of sizes and formats:
|CF® dye conjugates (13 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|
Secondary Antibodies – available with many conjugations
BUFFERS, FIXATIVES, OTHER ACCESSORIES
We offer the following accessory products for your flow cytometry experiments:
- 10X PBS, 4L
- Flow Cytometry Fixation/Permeabilization Kit
- Fixation Buffer
- Permeabilization Buffer
- Permeabilization and Blocking Buffer (5X)
- Mini Cell Scrapers
Click here to see all of our flow cytometry accessory products.