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LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit, for 405 nm excitation (Invitrogen™)

The LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit is used to determine the viability of cells prior to the fixation and permeabilization required for intracellular antibody staining or prior to elimination of biohazardous materials using formaldehyde fixation. This kit has been optimized and validated for use with a violet laser flow cytometer.

• Stable—dyes are freeze dried in separate vials to maintain stability
• Robust—staining pattern is the same before and after fixation
• Low compensation—minimal spectral overlap between other fluorophores

View a selection guide for all fixable viability dyes for flow cytometry.

Stable
Unlike products that are sold in solution, the LIVE/DEAD™ Fixable Aqua Stain has been conveniently packaged in 40-test vials to help ensure the stability and performance of the dye over time. Amine reactive dyes in solution will lose their effectiveness over a short period of time, therefore it is recommended to completely use the vial once rehydrated. If this is not possible, aliquot the vials in small volumes and store at -80°C, avoiding freeze-thaw cycles.

Robust
Dead cell discriminator stains can lose sensitivity after treatment with fixatives such as formaldehyde or ethanol-based fixation methods required for intracellular phosphorylation studies. The LIVE/DEAD™ Fixable Aqua Stain is an amine reactive dye that binds covalently to intracellular and extracellular amines, and the staining pattern is preserved following formaldehyde fixation.

Low compensation
The LIVE/DEAD™ Fixable Aqua Stain was selected based on its fluorescent properties to minimize compensation between other violet dyes and dyes that excite off of the 488 nm blue laser. The aqua-fluorescent reactive dye has an excitation maximum of ~375 nm, but it is excited well with the 405 nm violet laser, and it has an emission maxima of ~512 nm, so it can be collected in the second channel on most violet laser flow cytometers.

How it works
In cells with compromised membranes, the dye reacts with free amines both in the cell interior and on the cell surface, yielding intense fluorescent staining. In viable cells, the dye's reactivity is restricted to the cell-surface amines, resulting in less intense fluorescence. The difference in intensity is typically greater than 50-fold between live and dead cells, allowing for easy discrimination.

Colors available
LIVE/DEAD™ Fixable Dead Cell Stains are available in a wide variety of colors to meet your multi-color panel needs.

Amplex™ Red Monoamine Oxidase Assay Kit (Invitrogen™)

The Amplex Red Monoamine Oxidase Assay Kit provides a sensitive and simple fluorometric method for detecting monoamine oxidase activity in purified enzyme preparations at levels as low as 12 µU/mL and in tissue samples with as little as 20 µg protein in a 100 µL assay volume.

CellEvent™ Caspase-3/7 Green ReadyProbes™ Reagent (Invitrogen™)

CellEvent® Caspase-3/7 Green ReadyProbes® Reagent is a fluorogenic, no-wash indicator of activated caspase-3/7 for live- and fixed-cell applications. Activation of caspase-3 is an early indicator of apoptosis and CellEvent® Caspase-3/7 Green reagent allows rapid and sensitive detection of cells destined for cell death.

• Simple and fast no-wash protocol helps preserve delicate apoptotic cells
• Compatible with both live-cell fluorescence imaging and formaldehyde-based fixation methods
• Ready-to-use liquid formulation in convenient dropper bottle—no need to dilute, weigh, or pipette
• Stability at room temperature—keep handy at your scope or cell culture area.

See other ReadyProbes® reagents for cell staining
Learn more about other assays for apoptosis

Cell imaging applications
CellEvent® Caspase-3/7 Green reagent is a four amino acid peptide (DEVD) conjugated to a nucleic acid-binding dye that is non-fluorescent when not bound to DNA. The CellEvent® Caspase-3/7 Green reagent is intrinsically non-fluorescent, as the DEVD peptide inhibits binding of the dye to DNA. Upon activation of caspase-3/7 in apoptotic cells, the DEVD peptide is cleaved and the free dye can bind DNA, generating a bright green fluorescence. The fluorescence emission of the dye when bound to DNA is 530 nm and can be observed using a standard FITC filter set.

Suggestions for use
• In most cases, 2 drops/ml and an incubation time of 30 minutes is sufficient for bright nuclear staining of apoptotic cells; however, optimization may be needed for some cell types, conditions, and applications. In such cases, simply add more or fewer drops until the optimal staining intensity is obtained.
• If desired, combine CellEvent™ Caspase-3/7 Green ReadyProbes® reagent with a red or far-red nuclear stain for a total cell count
• CellEvent™ Caspase-3/7 Green dye is excited with a maximum at 502 nm and has an emission maximum at 530 nm. It is detected through standard GFP and FITC filters.

6-NBDG (6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-Deoxyglucose) (Invitrogen™)

6-NBDG is a fluorescent nonhydrolyzable glucose analog that has been used to monitor glucose uptake and transport in live cells. Although sensitive to its environment NBD fluorescence typically displays excitation/emission maxima of ~465/540 nm and can be visualized using optical filters designed for fluorescein.

eBioscience™ Human Regulatory T Cell Staining Kit #3 (Invitrogen™)

This Human Regulatory T cell Staining Kit contains all of the buffers and monoclonal antibodies for CD4, CD25, and Foxp3 necessary to successfully stain and identify regulatory T cells from human peripheral blood cells.

The RPA-T4 monoclonal antibody reacts with human CD4, a 59 kDa glycoprotein found on the surface of the majority of thymocytes, a subset of mature T cells (T helper cells), and at lower levels on monocytes. The BC96 monoclonal antibody reacts with human CD25 (also known as interleukin-2 receptor alpha, IL-2R alpha), a 55 kDa surface protein expressed by early progenitors of T cells and B cells, by mature, activated T cells and B cells, and at constitutively high levels on regulatory T cells. The PCH101 monoclonal antibody reacts with the amino terminus of human Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2. Foxp3 is a 49-55 kDa protein and a member of the forkhead/winged-helix family of transcription factors. It was identified as the gene responsible for the X-linked lymphoproliferative disease observed in scurfy (sf) mice and in the human disorder, X-linked autoimmunity-allergic dysregulation syndrome (XLAAD). Constitutive expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Reactivity/Species
Human

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Pierce™ LDH Cytotoxicity Assay Kit (Thermo Scientific™)

The Thermo Scientific Pierce LDH Cytotoxicity Assay Kit is a reliable colorimetric assay to quantitatively measure lactate dehydrogenase (LDH) released into the media from damaged cells as a biomarker for cellular cytotoxicity and cytolysis.

Features of the LDH Cytotoxicity Assay Kit:

Convenient—add-mix-read assay format for adherent and suspension cells
Colorimetric—quantitatively measures LDH release by formation of colored product
Robust—uses stable LDH enzyme activity as a cytotoxic marker
Flexible—ideal for high-throughput screening
Non-radioactive—safe alternative to 51Cr-release cytotoxicity assays

The Pierce LDH Cytotoxicity Assay Kit measures extracellular LDH in culture media using an enzymatic reaction that results in a red formazan product which can be measured spectrophotometrically. Lactate dehydrogenase (LDH) is a cytosolic enzyme that is is an indicator of cellular toxicity. The assay is ideal for high-throughput screening and provides a safe alternative to radioactive cytotoxicity assays.

Includes:
Kit contains substrate mix, assay buffer, 10X lysis buffer, stop solution, and LDH positive control

Requires:
Microplate reader capable of reading absorbance at 490nm and 680nm, flat-bottom clear 96-well plate compatible with spectrophotometer, multichannel pipette

Applications:
• Measure in vitro cytotoxicity mediated by chemicals, immune cells, or siRNA or microRNA
• Detect cytolysis in bioreactor or 3D tissue engineering applications

Lactate dehydrogenase (LDH) is a cytosolic enzyme present in many different types of cells. When the plasma membrane is damaged, LDH is released into cell culture media. The released LDH can be quantified by a coupled enzymatic reaction. First, LDH catalyzes the conversion of lactate to pyruvate via reduction of NAD+ to NADH. Second, diaphorase uses NADH to reduce a tetrazolium salt (INT) to a red formazan product. Therefore, the level of formazan formation is directly proportional to the amount of released LDH in the medium.

The assay is performed by transferring cell culture media from treated cells into a new microplate and adding the kit reagents. After incubation at room temperature for 30 minutes, reactions are stopped and LDH activity is determined by spectrophotometric absorbance at 490nm.

LIVE/DEAD™ Violet Viability/Vitality Kit, for 405 nm excitation (Invitrogen™)

The LIVE/DEAD™ Violet Viability/Vitality Kit provides a two-color fluorescence cell viability and vitality assay that is based on the simultaneous determination of live and dead cells with two probes that measure recognized parameters of cell health: plasma membrane integrity and intracellular esterase activity. Calcein violet AM and aqua-fluorescent reactive dye are optimal dyes for this application; both dyes utilize the violet laser allowing other laser lines to be used for conventional fluorochromes.

View a selection guide for all Nonfixable Viability Dyes for Flow Cytometry.

CaspGLOW™ Fluorescein Active Caspase-8 Staining Kit (Invitrogen™)

The CaspGLOW™ Fluorescein Active Caspase-8 Staining Kit contains all the reagents necessary to detect active caspase-8 in cells with high sensitivity. Fluorescein (FITC)-conjugated IETD-FMK, a specific inhibitor of caspase-8, is utilized in this assay for detection. This reagent is cell permeable, non-toxic and irreversibly binds to the active enzyme. Detection of the labeled cells can be determined by flow cytometry, fluorescent microscopy or a fluorescent plate reader.

The caspases constitute a family of aspartate-specific cysteine proteases that mediate a sequence of cleavage events. Recruitment of the inactive proenzyme to oligomerized receptors leads to caspase activation and autoproteolytic cleavage. These active enzymes can then cleave other caspases, thereby generating a caspase signaling cascade that leads to a form of programmed cell death termed apoptosis.

Caspase-8, which is also known as FLICE, MACHalpha1, and Mch5, cleaves its substrates at the C-terminal aspartic acid residue of the motif Asp-X-X-Asp. Eight isoforms of caspase-8 exist, of which caspase-8/a and 8/b are the predominant forms. Upon stimulation of death receptors such as CD95/APO-1/Fas, TRAIL-R1, TRAIL-R2, TNFR1, and TRAMP, caspase-8 is recruited to the death-inducing signaling complex (DISC). Subsequent dimerization leads to caspase-8 activation via autocatalytic cleavage, which leads to the formation of a 12-kDa prodomain and a 43-kDa intermediate fragment that is further cleaved to produce 26-kDa and 18-kDa active enzymes. As an initiator caspase, this enzyme initiates a caspase signaling cascade that results in apoptosis. Substrates of caspase-8 include caspases-3 and -7, as well as the pro-apoptotic Bcl-2 family member Bid. In addition to its role in cell death, caspase-8 has been linked to cell adhesion and motility.

Reported Application
Flow Cytometric Analysis, Immunocytochemistry

Dead Cell Apoptosis Kit with Annexin V Alexa Fluor™ 488 & Propidium Iodide (PI) (Invitrogen™)

The Dead Cell Apoptosis Kit with Annexin V Alexa Fluor™ 488 & Propidium Iodide is a flow cytometry kit used to measure early apoptosis by detecting phosphatidyl serine expression and membrane permeability.

View a selection guide for all apoptosis assays for flow cytometry.

Superior brightness
Unlike other Annexin V kits that have lower protein concentrations or purification levels, the Annexin V Alexa Fluor™ 488 conjugate is optimized for flow cytometry to provide the largest separation between apoptotic and live cells. The Alexa Fluor™ 488 dye is a superior green-fluorescent dye with a spectrum similar to fluorescein (FITC).

High binding efficiency
Annexin V conjugates are made from a highly purified cys-annexin, which leads to higher binding efficiency, resulting in highly accurate characterization of the apoptotic process.

Multi-parametric
Many publications require a minimum of two different ways to identify that cells are apoptotic. This multi-parametric kit detects phosphatidyl serine (PS) on the cytoplasmic surface of the cell membrane and membrane integrity using propidium iodide.

How it works
When cells are stained with Annexin V and propidium iodide, apoptotic cells expressing PS show green fluorescence, which can be detected in the FITC channel, and low red fluorescence. Dead or necrotic cells show bright red fluorescence and no green fluorescence, while live cells show no green or red fluorescence.

Arcturus™ Paradise™ PLUS qRT-PCR Kit (Applied Biosystems™)

The ARCTURUS® Paradise® PLUS Reagent System enables unprecedented gene expression analysis of millions of previously inaccessible samples. The Paradise® PLUS Reagent System is an integrated solution for gene expression studies from formalin-fixed, paraffin-embedded (FFPE) tissues, including reagents for tissue staining, RNA isolation and RNA amplification.

Key product features:
• Accelerated discovery process – unlock the tissue archives to access previously inaccessible samples
• Streamlined application – integrate seamlessly into current workflow
• Efficient workflow – secure downstream success with the Paradise® QC Kit
• Comprehensive system – benefit from the only complete system for formalin-fixed samples

Unlock the Tissue Archives
Currently, tens of millions of tissue samples with known outcomes are available in tissue archives. The Paradise® PLUS Reagent system enables microarray and qRT-PCR analysis of these previously inaccessible samples, allowing retrospective studies to accelerate discoveries. Achieve unprecedented results in just six steps (Figure 1).

Integrate Seamlessly into Current Workflow
The Paradise® PLUS Reagent System is compatible with all major gene expression platforms. Robust and reproducible results are achieved using tools and techniques commonly employed in today’s research laboratories.

Secure Downstream Success with the Paradise® QC Kit
Not all FFPE samples contain high quality RNA. Identify the best samples for your studies using a simple, real-time PCR research technique, and then proceed with confidence using these pre-qualified samples.

Benefit from the Only Complete System for Formalin-fixed Samples
The Paradise® PLUS Reagent System is the only available solution that provides all the components necessary to prepare RNA from formalin-fixed, paraffin-embedded tissue samples for gene expression analysis. To ensure successful gene expression results, the system includes reagents optimized for exceptional recovery of RNA and superior reverse transcription and RNA amplification (Figure 3).

Integrated Systems for Microgenomics
The Paradise® PLUS Reagent System is validated as part of Arcturus' complete Systems for Microgenomics®, an integrated solution for utilizing small quantities of RNA for gene expression analysis, and features the ARCTURUSXT Laser Capture Microdissection (LCM) Instrument to procure pure cell populations. ARCTURUS®' Systems for Microgenomics enable accurate and sensitive microarray assays that reveal molecules otherwise obscured in whole tissue samples or cell mixtures.

For Research Use Only. Not for use in diagnostics procedures.

LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy & quantitative assays (Invitrogen™)

The LIVE/DEAD BacLight Bacterial Viability Kit *for microscopy and quantitative assays* is a convenient and easy-to-use kit for monitoring the viability of bacterial populations as a function of the membrane integrity of the cell. Cells with a compromised membrane that are considered to be dead or dying will stain red, whereas cells with an intact membrane will stain green.

View additional information about all microbiological analysis products.

eBioscience™ Mouse Hematopoietic Lineage Biotin Panel (Invitrogen™)

The eBioscience Mouse Hematopoietic Lineage Flow Panel contains 5 biotinylated antibodies that can be used to identify, enrich and/or deplete cells committed to the T, B, NK, myeloid and erythroid lineages based on their cell surface antigen expression.

Reactivity/Species
Mouse

Reported Application
Flow Cytometric Analysis

eBioscience™ Human Regulatory T Cell Whole Blood Staining Kit (Invitrogen™)

This Human Regulatory T Cell Whole Blood Staining Kit contains the buffers and monoclonal antibody necessary to successfully stain and identify Foxp3 cells in whole blood samples. The PCH101 monoclonal antibody reacts with the amino terminus of human Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2. Foxp3 is a 49-55 kDa protein and a member of the forkhead/winged-helix family of transcription factors. It was identified as the gene responsible for the X-linked lymphoproliferative disease observed in scurfy (sf) mice and in the human disorder, X-linked autoimmunity-allergic dysregulation syndrome (XLAAD). Constitutive expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

The PCH101 antibody crossreacts with rhesus, chimpanzee, and cynomolgus Foxp3 PCH101 recognizes a different epitope of Foxp3 than clones 236A/E7 and 150D/E4.

Host
Rat

Isotype
IgG2a, kappa

Reactivity/Species
Human

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Escherichia coli (K-12 strain) BioParticles™, Alexa Fluor™ 594 conjugate (Invitrogen™)

The Molecular Probes® BioParticles® product line consists of a series of fluorescently labeled, heat- or chemically killed bacteria and yeast in a variety of sizes, shapes, and natural antigenicities. These fluorescent BioParticles® products have been employed to study phagocytosis by fluorescence microscopy, quantitative spectrofluorometry, and flow cytometry.

We offer E. coli (K-12 strain), S. aureus (Wood strain, without protein A) and zymosan (S. cerevisiae) BioParticles® conjugates covalently labeled with a variety of different fluorophores (special care has been taken to remove free dye after conjugation). Unlike the fluorescence of fluorescein-labeled BioParticles® conjugates, which is partially quenched in acidic environments, the fluorescence of the Alexa Fluor®, BODIPY® FL, tetramethylrhodamine and Texas Red® dye conjugates is uniformly intense over the pH range from 3 to 10.

BioParticles Specifications:
• Label (Ex/Em): Alexa Fluor® 594 (~590/617 nm)
• Particle: E. coli (K-12 strain)
Opsonizing reagent available


Using BioParticles Products
BioParticles® conjugates are provided as lyophilized powders. There are approximately 3 x 108 E. coli or S. aureus particles per mg solid and approximately 2 x 107 zymosan particles per mg solid. BioParticles® conjugates can be reconstituted in the buffer of your choice for use in phagocytosis assays. The fluorescence of BioParticles® conjugates that are bound to the surface of the cell (but not internalized) can be quenched by ethidium bromide, trypan blue, or other quenchers. In addition to cellular applications, fluorescent BioParticles® conjugates may be effective as flow cytometry calibration references when sorting bacteria and yeast mutants. These small particles may also be useful references for light scattering studies because their sizes and shapes differ in characteristic ways.

Find More BioParticles® Products
We offer a large range of dye-labeled and unlabeled E. coli (K-12 strain), S. aureus (Wood strain, without protein A), and zymosan (S. cerevisiae) BioParticles® products. Find out about these products and their applications by reviewing Probes for Following Receptor Binding and Phagocytosis—Section 16.1 in the Molecular Probes® Handbook.

For pH-sensitive endocytosis assays, see our pHrodo™ BioParticles® conjugates.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

PrestoBlue™ HS Cell Viability Reagent (Invitrogen™)

PrestoBlue HS Cell Viability Reagent is a ready-to-use resazurin-based solution that functions as a cell health indicator by using the reducing power of living cells to quantitatively measure viability. Upon entering living cells, resazurin is reduced to resorufin, a compound that is red in color and highly fluorescent. High background fluorescence caused by various contaminants reduces the performance of resazurin-based reagents. To improve the performance an innovative purification process was developed that removes these contaminants. This highly purified, non-toxic resazurin was used in the standard PrestoBlue formulation, creating the PrestoBlue HS Cell Viability Reagent. Changes in viability can be easily detected using either an absorbance- or fluorescence-based plate reader. The PrestoBlue HS Cell Viability Reagent has been optimized for the fast and reliable detection of mammalian cell viability.

PrestoBlue HS Cell Viability Reagent features include:
• Removal of contaminants from resazurin—displays a >50% reduction in background fluorescence
• Signal-to-background ratio increased by >100%—results in large assay signal window
• Highly sensitive reagent with a linear response—detects as few as 10 cells per well
• Convenient add-and-read format—no mixing, no washing, no cell lysis, and compatible with either fluorescence- or absorbance-based instrumentation
• Optimized for reliable detection of mammalian cell viability in only 10 minutes

Measuring changes in cell viability is a fundamental method for assessing cell health, determining genotoxicity, and evaluating anti-cancer drugs. Cell health can be monitored by detecting changes in several key indicators, including changes to plasma membrane integrity, DNA synthesis, DNA content, enzyme activity, presence of ATP, and cellular reducing environment.

Monitoring changes to the cellular reducing environment or metabolic activity by using resazurin-based reagents is a well-established and reliable indicator of cell viability or death. Upon entering living cells, the cellular reducing environment reduces resazurin to resorufin, a compound that is red in color and highly fluorescent. Viable cells continuously convert resazurin to resorufin, increasing the overall fluorescence of the medium surrounding the cells. Additionally, the conversion of resazurin to resorufin results in a pronounced color change, thus cell viability can also be detected using absorbance-based plate readers.

As a consequence of the synthesis and manufacturing processes, all resazurin-based reagents contain a detectable amount of resorufin contamination. The amount of contamination can vary greatly between sources of the material and manufacturing conditions, contributing to differences in detectable background fluorescence. An innovative process was developed that removes the contaminating resorufin, resulting in the highly pure resazurin used in PrestoBlue HS Cell Viability Reagent. This highly pure resazurin produces a >50% decrease in background fluorescence and a >100% increase in the signal-to-background ratio. Mammalian cell viability can be detected after a 10-minute incubation with PrestoBlue HS Cell Viability Reagent. Since no lysis is required, the diluted PrestoBlue HS solution can be removed and replaced with complete growth medium, for further culturing of the cells.