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pHrodo™ Red S. aureus Bioparticles™ Conjugate for Phagocytosis (Invitrogen™)

New pH-sensitive pHrodo® Red dye conjugates give faster and more accurate results than any other phagocytosis assay pHrodo® Red dye conjugates are non-fluorescent outside the cell, but fluoresce brightly red in phagosomes.

Get faster staining and more accurate results without the need for wash steps or quencher dye
• Specific detection of phagocytosis and endocytosis
• Reduced signal variability and improved timing in sensitive experiments
• Multiplex with green dyes such as GFP, Fluo-4, or calcein

The fluorescence of the novel pHrodo® Red dye dramatically increases as pH decreases from neutral to the acidic, making it an ideal tool to study phagocytosis and its regulation by drugs and/or environmental factors. The lack of fluorescence outside the cell eliminates the need for wash steps and quencher dyes. Use the ready-made pHrodo® Red S. aureus BioParticles® conjugates in imaging or flow applications, or pHrodo® Red SE, the activated succinimidyl ester, for labeling microorganisms or proteins of your choice.

CellEvent™ Caspase-3/7 Green Flow Cytometry Assay Kit (Invitrogen™)

The CellEvent™ Caspase-3/7 Green Flow Cytometry Assay Kit enables the flow cytometric detection of activated caspase-3 and caspase-7 in apoptotic cells. The kit includes the novel fluorogenic substrate CellEvent™ Caspase-3/7 Green Detection Reagent, as well as SYTOX™ AADvanced™ Dead Cell Stain.

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

• Caspase-3/7 specific—includes the recognition sequence for activated caspase-3 and caspase-7
• Easy identification—clearly identify live, dead, and apoptotic cell populations
• Quick analysis—washing and fixation is not required
• Multicolor compatibility—combine with other dyes excitable by the 488 nm laser or other lasers

CellEvent Caspase-3/7 Green Detection Reagent is a cell-permeant reagent that consists of a four-amino acid peptide (DEVD, a caspase 3/7 recognition sequence) conjugated to a nucleic acid-binding dye. During apoptosis, caspase-3 and caspase-7 proteins are activated and cleave the peptide. Cleavage of the peptide allows binding of DNA by the reagent, thereby labeling the apoptotic cells with a bright, fluorogenic signal that has absorption/emission maxima of ~502/530 nm. When used together with the SYTOX AADvanced Dead Cell Stain, apoptotic cells can be easily discriminated from live and necrotic cells.

Because no single parameter defines apoptosis in all systems, we strongly suggest using a combination of different measurements for reliable detection of apoptosis. For more information, see Apoptosis Reagents in Flow Cytometry.

CaspGLOW™ Fluorescein Active Caspase-9 Staining Kit (Invitrogen™)

The CaspGLOW™ Fluorescein Active Caspase-9 Staining Kit contains all the reagents necessary to detect active caspase-9 in mammalian cells with high sensitivity. Fluorescein (FITC)-conjugated LEHD-FMK, a specific inhibitor of caspase-9, 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-9, which is also known as ICE-LAP6 and Mch6, cleaves its substrates at the C-terminal aspartic acid residue of the motif Asp-X-X-Asp. Activation of this signaling protease occurs upon stimulation of death receptors such as CD95/APO-1/Fas, TRAIL-R1, TRAIL-R2, TNFR1, and TRAMP. Caspase-9 is activated by recruitment and dimerization within the Apaf-1 apoptosome. Once recruited, caspase-9 undergoes proteolytic cleavage at Asp315 to yield 35-kDa and 10-kDa fragments. Unlike other caspases, this cleavage event is not required for the catalytic activity of caspase-9. As an initiator caspase, this protease initiates a caspase signaling cascade that results in apoptosis. Substrates of caspase-9 include caspases-3, -6, and -7. Finally, dimerization of cleaved caspase-9 is inhibited by X-linked inhibitor of apoptosis (XIAP).

Reported Application
Flow Cytometric Analysis, Immunocytochemistry

eBioscience™ BrdU Staining Kit for Flow Cytometry APC (Invitrogen™)

The eBioscience BrdU Staining Kit for Flow Cytometry contains the necessary reagents and buffers for identifying and examining proliferating cells of mammalian species by flow cytometric analysis. Cycling cells are incubated with 5-bromo-2’-deoxyuridine (BrdU), a synthetic analog of thymidine which incorporates into newly synthesized genomic DNA during the S-phase of mitosis. Following DNA denaturation, the cells are stained for BrdU incorporation along with any other cell surface and/or intracellular targets of interest.

This kit is optimized to achieve brighter staining compared to traditional protocols.

Reactivity/Species
Bovine, Canine, Cow, Human, Monkey, Mouse, Non-human primate, Rat, Rhesus

Conjugate
APC

Laser
Red Laser

Emit
660 nm

Excite
633 - 647 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Arcturus™ Paradise™ PLUS 2 Round Kit, biotin labeling, no solvents (Applied Biosystems™)

The ARCTURUS® Paradise® PLUS Reagent System with Microarray Labeling permits 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. Amplify RNA from as little as 5 ng of fixed RNA using methods that have been specially optimized to overcome the challenges of cross-linked templates.

Key product features:
• Microarray Labeling, Gene Expression Profiling
• Flexibility in Assay Format
• Integrated Systems for Microgenomics

Microarray Labeling, Gene Expression Profiling
The Paradise® PLUS RNA Amplification Kit may be configured to include Turbo Labeling™ for use in microarray experiments. Turbo Labeling Kits offer a proprietary, non-enzymatic technology optimized for the labeling of unmodified, amplified RNA (aRNA) for gene expression profiling. The unmodified aRNA is labeled post-amplification, thereby avoiding the need to incorporate modified nucleotides during RNA amplification. The use of unmodified nucleotides in the amplification process results in unmodified aRNA with higher yields and longer aRNA fragments, thus providing better representation of the mRNA transcript for downstream analysis.

Flexibility in Assay Format
Turbo Labeling Kits include reagents to support labeling of 12 samples using either Cy™3⁄Cy™5 dyes or Biotin for hybridization to cDNA or oligonucleotide arrays.

Integrated Systems for Microgenomics
The Paradise® PLUS Reagent System is validated as part of the ARCTURUS® Complete System 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 molecular otherwise obscured in whole tissue samples or cell mixtures.

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

ELF™ 97 Immunohistochemistry Kit (Invitrogen™)

The ELF® 97 Immunohistochemistry Kit can be used to detection biotinylated probes including secondary antibodies. The kit also includes the ELF® 97 phosphatase substrate that upon hydrolysis produces a bright and photostable yellow-green fluorescent precipitate at the site of enzymatic activity. This fluorescent precipitate has several unique spectral characteristics, including an extremely large Stokes shift, 180 nm that makes it easily distinguishable from endogenous fluorescence.

HCS DNA Damage Kit (Invitrogen™)

The HCS DNA Damage kit simultaneously and quantitatively measures two important cell-health parameters: DNA damage and cytotoxicity. DNA damage is detected using an antibody against phosphorylated H2AX (Ser139) which is induced in response to double-strand break (DSB) formation. To detect cytotoxicity, the Image-iT® DEAD™ Green is an impermeant dye in healthy cells that becomes permeant when the plasma membrane of cells is compromised. Both indicators are amenable to fixation and permeabilization, thus allowing for multiplexing with other biomarkers of interest. The blue-fluorescent nuclear segmentation tool Hoechst 33342 is also included and stains DNA in both live and dead cells.

LIVE/DEAD™ FungaLight™ Yeast Viability Kit, for flow cytometry (Invitrogen™)

The LIVE/DEAD® FungaLight™ Yeast Viability Kit enables researchers to easily, reliably and quantitatively distinguish live and dead yeast in minutes by flow cytometry. The kit contains solutions of the SYTO® 9 green-fluorescent nucleic acid stain and the red-fluorescent nucleic acid stain, propidium iodide. These stains differ both in their spectral characteristics and in their ability to penetrate healthy yeast cells. When used alone, the SYTO® 9 stain generally labels all yeast in a population - those with intact membranes and those with damaged membranes. In contrast, propidium iodide penetrates only yeast with damaged membranes, causing a reduction in the SYTO® 9 stain fluorescence by fluorescence resonance energy transfer (FRET) when both dyes are present. As a result, yeast with intact membranes stain fluorescent green, whereas yeast with damaged membranes stain fluorescent red.

LIVE/DEAD™ Reduced Biohazard Cell Viability Kit #1, green & red fluorescence (Invitrogen™)

The LIVE/DEAD Reduced Biohazard Cell Viability Kit #1 is a convenient and easy-to-use cell viability kit that is designed to reduce the risk associated with handling potential biohazards such as viral, bacterial or protozoan pathogens. The assay monitors viability 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.

pHrodo™ Red Zymosan Bioparticles™ Conjugate for Phagocytosis (Invitrogen™)

New pH-sensitive pHrodo® Red dye conjugates give faster and more accurate results than any other phagocytosis assay. pHrodo® Red dye conjugates are non-fluorescent outside the cell, but fluoresce brightly red in phagosomes. Get faster staining and more accurate results—without the need for wash steps or quencher dye.

• Specific detection of phagocytosis and endocytosis
• Reduced signal variability and improved timing in sensitive experiments
• Multiplex with green dyes such as GFP, Fluo-4, or calcein

The fluorescence of the novel pHrodo® Red dye dramatically increases as pH decreases from neutral to acidic, making it an ideal tool to study phagocytosis and its regulation by drugs and/or environmental factors. The lack of fluorescence outside the cell eliminates the need for wash steps and quencher dyes.

Use the ready-made pHrodo® Red Zymosan BioParticles® Conjugate in imaging or flow applications, or pHrodo® Red SE, the activated succinimidyl ester, for labeling microorganisms or proteins of your choice.

In addition, pHrodo® dye is also available in green color, with E. coli, zymosan, S. aureus, and dextran 10,000 conjugates.

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

EnzChek™ Caspase-3 Assay Kit #1, Z-DEVD-AMC substrate (Invitrogen™)

The EnzChek Caspase-3 Assay Kit #1 allows detection of apoptosis by providing a simple and reliable method for assaying caspase-3 activity. The basis for the assay is the aminomethylcoumarin (AMC)-derived substrate Z-DEVD-AMC. This substrate, which is weakly fluorescent in the UV range (excitation/emission maxima ~330/390 nm), yields a bright, blue-fluorescent product (excitation/emission maxima ~342/441 nm) upon proteolytic cleavage. The kit can be used to continuously measure the activity of caspase-3 and closely related proteases in cell extracts and purified enzyme preparations, using a fluorometer or fluorescence microplate reader.

Dead Cell Apoptosis Kit with Annexin V APC and SYTOX™ Green, for flow cytometry (Invitrogen™)

This flow cytometry product detects the externalization of phosphatidylserine in apoptotic cells using recombinant annexin V conjugated to red laser-excited allophycocyanin, and dead cells using SYTOX™ Green nucleic acid stain. Apoptotic cells are detected by annexin V binding to externalized phosphatidylserine, and late apoptotic and necrotic cells have compromised membranes that permit SYTOX™ Green stain access to cellular nucleic acids. After exposing a cell population to APC annexin V and SYTOX™ Green stain, live cells show little or no fluorescence, apoptotic cells show red fluorescence and very little green fluorescence, and late apoptotic cells show a higher level of red and orange fluorescence. These populations can easily be distinguished using a flow cytometer that has both 488 nm and 633 nm excitation sources (an argon-ion laser and a HeNe laser).

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

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

2-NBDG is a fluorescent glucose analog that has been used to monitor glucose uptake in live cells, as an indicator of cell viability. 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.

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.

Click-iT™ EdU Alexa Fluor™ 647 HCS Assay (Invitrogen™)

The Click-iT® EdU Alexa Fluor® 647 HCS Assay is a superior alternative to traditional proliferation assays that is optimized for high-content imaging applications. In this assay the modified thymidine analogue EdU is efficiently incorporated into newly synthesized DNA and fluorescently labeled with a bright, photostable Alexa Fluor® dye in a fast, highly-specific click reaction. This fluorescent labeling of proliferating cells is accurate and compatible with antibody methods due to the mild click protocol.

• Simple—works the first time, every time, in less time
• Efficient—no denaturation steps or harsh treatment required
• Content-rich results—better preservation of cell morphology, antigen structure, and DNA integrity
• Consistent—not dependent on variable antibody lots for detection

The most accurate proliferation-detection methods are based on the incorporation and measurement of nucleoside analogues in newly synthesized DNA, with bromodeoxyuridine (BrdU) a commonly used analogue. BrdU-labeled DNA is quantitated using anti-BrdU antibodies following DNA denaturation by harsh methods (HCl, heat, or enzymes) to expose the BrdU molecules. This step is time consuming and difficult to perform consistently. The harsh treatment can also adversely effect sample integrity and quality, which makes co-staining with other antibodies challenging.

Superior Proliferation Methodology
The Click-iT® EdU Alexa Fluor® 647 HCS Assay provides a superior alternative to BrdU assays for measuring cell proliferation. EdU (5-ethynyl-2'-deoxyuridine) is a nucleoside analog of thymidine and is incorporated into DNA during active DNA synthesis. With Click-iT® EdU, mild fixation and detergent permeabilization is sufficient for the small molecule-based Click-iT® EdU detection reagent to gain access to the DNA. As a consequence, the Click-iT® EdU HCS assay is not only easy to use, but more accurate and compatible with cell cycle analysis and other intracellular or extracellular targets for truly content-rich results.

The kit is optimized for high-content imaging applications; visit the Click-iT® technology area of our website for kits designed for fluorescence microscopy, microplate, or flow cytometry platforms.

Learn more about Click-iT® technology

Notes:
The Click-iT® assay can be used on cells in culture or in vivo following administration of EdU by feeding or injection methods.
The Click-iT® assay can be used with BrdU in dual pulse experiments by using the ant-BrdU (clone MoBu-1) antibody, which does not cross react with EdU.
The Click-iT® technology is compatible with immunohistochemical, immunocytochemical, and fluorescent dyes that are fixation tolerant or designed for fixed-cell labeling.