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CyQUANT™ Cell Proliferation Assay, for cells in culture (Invitrogen™)

The CyQUANT® Cell Proliferation Assay is a highly sensitive fluorescence-based method for quantifying cells and assessing cell proliferation and cytotoxicity. For cell proliferation assays that are more compatible with high throughput screening, see our CyQUANT® NF Cell Proliferation Assay or our CyQUANT® Direct Assay.

With the CyQUANT® Cell Proliferation Assay, you can:

• Measure DNA content to directly quantify 50 to 50,000 cells per well without relying on metabolic activity
• Quickly compare multiple samples in a simple plate-based assay format
• Freeze and store time course samples for later analysis

Measure DNA content directly
The main component of the CyQUANT® Cell Proliferation Assay is CyQUANT® GR, a proprietary dye that exhibits strong fluorescence enhancement when bound to nucleic acids. With this dye, you can measure a sample's DNA content and compare it to standards, directly quantifying the entire cell population within a broad linear detection range. This method offers improved accuracy over metabolically based cell proliferation or cytotoxicity assays that can be influenced by cell changes that are unrelated to cell number.

Quickly compare multiple samples
The CyQUANT® Cell Proliferation Assay requires minimal hands-on time. Simply remove the culture media, freeze and lyse cells, then read fluorescence on a standard microplate reader with a fluorescein filter.

Store samples for later analysis
Because the cells are frozen, samples can stored for up to 4 weeks and run in batches, allowing the direct comparison of samples taken at different time points.

Kit components
The kit includes CyQUANT® GR dye and cell lysis buffer sufficient to perform 1,000 assays in volumes suitable for fluorescence detection in microplates. The kit also contains bacteriophage λ DNA to be used as a reference standard for assay calibration.

FUN™ 1 Cell Stain (Invitrogen™)

FUN® 1 is a unique two-color fluorescent viability probe for yeast and fungi. The FUN® 1 stain passively diffuses into a variety of cell types and initially stains the cytoplasm with a diffusely distributed green fluorescence. However, in several common species of yeast and fungi, subsequent processing of the dye by live cells results in the formation of distinct vacuolar structures with compact form that exhibit a striking red fluorescence, accompanied by a reduction in the green cytoplasmic fluorescence. Formation of the intravacuolar structures requires both plasma membrane integrity and metabolic capability. Dead cells fluoresce bright yellow-green, with no discernable red structures.

HCS LipidTOX™ Red Phospholipidosis Detection Reagent (1000X), for cellular imaging (Invitrogen™)

The HCS LipidTOX™ Red phospholipidosis detection reagent was developed to characterize the potentially toxic side effects of compounds on lipid metabolism in mammalian cell lines. This reagent is easily detected by fluorescence microscopy or an HCS reader equipped with filters that can detect excitation/emission maxima ~595/615 nm. LipidTOX™ phospholipidosis detection reagents do not affect the normal growth of cells, and the staining observed after incubation in live cells is maintained after formaldehyde fixation. This probe is compatible with the HCS LipidTOX™ neutral lipid stains (H34475, H34476, and H34477).

Arcturus™ Paradise™ PLUS 2 Round Kit, amino allyl (Applied Biosystems™)

The ARCTURUS® Paradise® PLUS Reagent System permits unprecedented gene expression analysis of millions of previously inaccessible samples. It 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.

This kit provides reagents for 2-round in-vitro transcription with aminoallyl aRNA labeling.

Key product features:
• Accelerated discovery process – unlock the tissue archives to access previously inaccessible samples
• Simplified procedure – achieve unprecedented results in six steps
• 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 Six Steps
Only six steps are required to take you from sample to results (See Figure 1).

Integrate Seamlessly into Current Workflow
The Paradise® PLUS Reagent System is compatible with all major microarray platforms. Robust and reproducible results are achieved using tools and techniques commonly employed in today’s research laboratories (See Figures 2, 3, 4, and 5).

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 (See Figure 6).

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 microarray results, the system includes reagents optimized for exceptional recovery of RNA and superior RNA amplification.

Integrated Systems for Microgenomics
The Paradise® PLUS Reagent System is a validated as part of the Arcturus® Complete System for Microgenomics®, an integrated solution for utilizing small quantities of RNA for gene expression analysis. This system 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 signatures otherwise obscured in whole tissue samples or cell mixtures.

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

CyQUANT™ LDH Cytotoxicity Assay (Invitrogen™)

The CyQUANT LDH Cytotoxicity Assay is a colorimetric assay that provides a simple and reliable method for determining cellular cytotoxicity. Lactate dehydrogenase (LDH) is a cytosolic enzyme present in many different cell types that is released into the cell culture medium upon damage to the plasma membrane. The CyQUANT LDH Cytotoxicity Assay provides the reagents to accurately and quantitatively measure this extracellular LDH.

Note: The CyQUANT LDH Cytotoxicity Assay (Cat. Nos. C20300 and C20301) are direct replacements for the Pierce LDH Cytotoxicity Assay Kit (Cat. Nos. 88953 and 88954).

CyQUANT LDH Cytotoxicity Assay features include:
Convenient—add-mix-read assay format for adherent and suspension cells, including 3D cell models
Accurate—provides a quantitative measurement of LDH release
Flexible—ideal for high-throughput screening, monitor cytotoxicity from the same sample over time
Robust—uses stable LDH enzyme activity as a cytotoxic marker

LDH is a cytosolic enzyme present in many different cell types and is a well-established and reliable indicator of cellular toxicity. Damage of the plasma membrane results in a release of LDH into the surrounding cell culture medium. This extracellular LDH can be quantified by a coupled enzymatic reaction in which LDH catalyzes the conversion of lactate to pyruvate via NAD+ reduction to NADH. Diaphorase then uses NADH to reduce a tetrazolium salt (INT) to a red formazan product that can be measured at 490 nm. The level of formazan formation is directly proportional to the amount of LDH released into the medium.

The CyQUANT LDH Cytotoxicity Assay provides the reagents needed for the simple, reliable colorimetric quantification of cellular cytotoxicity. The kit can be used with different cell types, including 3D cell models, to measure cytotoxicity mediated by chemical compounds as well as cell-mediated cytotoxicity. Since LDH in the medium is the indicator of cellular cytotoxicity, the assay can be used to monitor cytotoxicity from the same sample over time. To perform the assay, an aliquot of the cell culture medium is transferred to a new plate and the reaction mixture is added. After a 30-minute incubation, the reaction is stopped by adding Stop Solution and absorbance is measured using a microplate reader.

pHrodo™ Green E. coli BioParticles™ Conjugate for Phagocytosis (Invitrogen™)

New pH-sensitive pHrodo® Green dye conjugates, like our pHrodo® Red dye conjugates, give faster and more accurate results than any other phagocytosis assay. pHrodo® Green conjugates are non-fluorescent outside the cell at neutral pH, but fluoresce brightly green at acidic pH such as 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
• Enough to do 100 tests when using 100 µL in each well of 96-well plate
• Multiplex with other compatible dyes, such as red florescent protein, TMRM, NucBlue™ Hoechst, and CellROX® Deep Red

The fluorescence of the novel pHrodo® Green 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® Green E. coli BioParticles® Conjugate in imaging, high content screening, high throughput screening, and flow applications. pHrodo® Green dye is also available as a conjugate of zymosan BioParticles®, S. aureus BioParticles®, or dextran 10,000 MW. To create other conjugates, such as antibody conjugates, use pHrodo® Green STP ester or pHrodo® Green maleimide.

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

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

This product detects the externalization of phosphatidylserine in apoptotic cells using recombinant annexin V conjugated to the orange fluorescent phycobiliprotein R-PE, and dead cells using SYTOX™ Green nucleic acid stain. After treatment with both probes, apoptotic cells show orange fluorescence, dead cells show green fluorescence, and live cells show little or no fluorescence. These populations can easily be distinguished in the 530/30 nm and 585/42 nm bandpass filters with a 488 nm laser flow cytometer.

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

eBioscience™ Essential Human T-Cell Phenotyping Kit (Invitrogen™)

The Invitrogen eBioscience Essential Human T-Cell Phenotyping Kit is designed for flow cytometry-based identity testing of T-cells. It leverages high-quality eBioscience antibodies and isotype controls and includes a ready-to-use protocol for experimental setup, gating strategy, and analysis.

Benefits include:
Proven performance―antibodies and supporting reagents designed for flow cytometry
Highly referenced antibodies―eBioscience portfolio contains trusted fluorochrome-conjugated antibodies and reagents for immunology and oncology research
Turnkey kit and protocol―contains reagents and standardized gating recommendations needed to perform flow cytometry characterization

Arcturus™ Paradise™ PLUS Whole Transcript Reverse Transcription Kit (WT-RT) (Applied Biosystems™)

The ARCTURUS® Paradise® PLUS Whole Transcript RT (WT-RT) Reagent System provides an efficient and reliable solution for purifying RNA from archived Formalin-Fixed Paraffin-Embedded (FFPE) tissue samples followed by reverse transcription of whole transcript for use in real-time PCR.

FFPE samples introduce unique challenges for gene expression profiling and gene expression quantitation, including chemical modification and fragmentation of RNA molecules. Storage of FFPE samples adds to these challenges through increased RNA degradation over time.

The Paradise® PLUS WT-RT Reagent System was specifically developed and specially optimized to provide high-quality RNA and robust reverse transcription (RT) from highly fragmented RNA obtained from archived FFPE samples over 20 years old (Figure 2).

Key product features:
• Simple workflow—minimal sample handling reduces chance of contamination (Figure 3)
• Efficient RNA extraction and isolation—saves time and minimizes sample loss
• Optimized RT reactions—generate cDNA representing the entire length of the transcript
• Accurate transcript analysis—results represent high, medium, and low-abundance expressing genes

Optimized RNA Extraction and Isolation from Archived FFPE Tissues
Figure 1 shows a comparison between total RNA yields using the Paradise® PLUS WT-RT Reagent System and a similar commercially available protocol. The average yield of the three replicate isolations illustrates the abundance of total RNA made available using the Paradise® PLUS WT-RT Reagent System. The Paradise® PLUS WT-RT Reagent System produces abundant total RNA for reverse transcription and subsequent qPCR analysis, up to three times more total RNA than obtained when a similar commercially available product was used for the same sample.

Superior Amplification Possible Over Wide Range of Targets
The sensitivity of the Paradise® PLUS WT-RT Reagent System was compared to similar reagent products available on the market that generate cDNA for use in real-time PCR reactions. As Figure 4 illustrates, improved Ct values were seen for all high-, medium- and low-expressing genes when the Paradise® PLUS product was used. Exon-spanning primers at varying distances from the 3’-end of the transcript (500 bp to 5 kb) were used to show increased sensitivity and improved transcription success compared to other commercially available products. The Paradise® PLUS WT-RT Reagent System successfully amplified all genes, suggesting the reagents provided in the kit are optimized to work together, saving you time and effort while producing high-quality results across the entire transcript.

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 permit 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.

CellROX™ Green Flow Cytometry Assay Kit (Invitrogen™)

The CellROX™ Green Flow Cytometry Assay Kit enables the flow cytometric detection of reactive oxygen species (ROS) in live cells. The kit includes the novel, fluorogenic CellROX™ Green Reagent, as well as SYTOX™ Red Dead Cell Stain, N-acetyl cysteine (an antioxidant, for negative control), and tert-butyl hydroperoxide solution (TBHP, an inducer of ROS).

View a selection guide for all CellROX® Reagents and Kits.

The CellROX™ Green Flow Cytometry Assay Kit features:

• Fluorogenic probe formulated for flow cytometry that is oxidized in the presence of ROS
• Multicolor compatibility—minimal overlap with fluorophores excited by other laser lines, allowing easy multiplexing with other reagents
• Simple protocol—cells can be stained in complete media or other appropriate buffer, no need for serum free media

The CellROX™ Green Detection Reagent is cell-permeable and non-fluorescent or very weakly fluorescent while in the reduced state. Upon oxidation, the reagent binds DNA and exhibits a strong fluorogenic signal that has absorption/emission maxima of 508/525 nm and remains localized to the nucleus and cytoplasm. When used together with the included SYTOX™ Red Dead Cell Stain, oxidatively stressed and nonstressed cells are reliably distinguished from dead cells by flow cytometry.

eBioscience™ Essential Human T-Cell Phenotyping Panel (Invitrogen™)

The Invitrogen eBioscience Essential Human T-Cell Phenotyping Panel is designed for flow cytometry-based identity testing of T-cells. It leverages high quality eBioscience antibodies and a ready-to-use protocol for experimental setup, gating strategy, and analysis.

Benefits include:
Proven performance―antibodies designed for flow cytometry
Highly referenced antibodies―eBioscience portfolio contains trusted fluorochrome-conjugated antibodies for immunology and oncology research
Turnkey kit and protocol―contains standardized gating recommendations needed to perform flow cytometry characterization

CellTrace™ Violet Cell Proliferation Kit, for flow cytometry (Invitrogen™)

CellTrace™ Violet Cell Proliferation Kit is used for in vitro and in vivo labeling of cells to trace multiple generations using dye dilution by flow cytometry.

• Superior performance—bright, single-peak staining enables visualization of multiple generations
• Long-term signal stability—well-retained in cells for several days post stain
• Versatile—multiple colors available to easily combine with antibodies or markers of cell function, such as GFP
• Simple, robust staining protocol

View a selection guide for all CellTrace™ Cell Proliferation Kits for flow cytometry.

Superior fluorescent staining
Successful proliferation analysis by dye dilution (see figure below) requires an extremely bright dye to distinguish fluorescently labeled cells from auto-fluorescence after several cell divisions. The intense fluorescent staining provided by CellTrace™ Violet dye enables the visualization of ten or more generations of proliferating cells before the signal is overwhelmed by intrinsic cellular auto-fluorescence. Consistent, homogeneous staining results in very little fluorescence variation between cells in a population, so distinct generations can be seen without any requirement for complex modeling software.

Long-term signal retention
Unlike stains that label the lipid membrane of cells, CellTrace™ Violet stain easily crosses the plasma membrane and covalently binds inside cells where the stable, well-retained fluorescent dye provides a consistent signal, even after several days in a cell culture environment. CellTrace™ Violet dye binds covalently to all free amines on the surface and inside of cells and shows little cytotoxicity, with minimal observed effect on the proliferative ability or biology of cells.

Easy multiplexing with other fluorophores
The violet excitation and narrow emission of CellTrace™ Violet dye make it ideal for multiplexing due to the limited spectral overlap with other common dyes (Alexa Fluor™ 488, FITC, and RPE) and fluorescent proteins (Green Fluorescent Protein (GFP) and mCherry) (see Fluorescence Spectra for CellTrace™ Violet stain below).

Simple, robust staining protocol
The CellTrace™ Violet Cell Proliferation Kit contains convenient single-use vials of dry dye to permit small-scale experiments without preparing excess quantities of dye. A stock solution is prepared by dissolving the contents of a vial in anhydrous DMSO prior to use. To stain 1 mL of cells in protein-free medium, 1 µL of this stock solution is typically used. Cells should be stained for 20 minutes at room temperature with gentle agitation. A brief wash with complete medium will then quench any dye remaining in solution.

MDR1 Vesicular Transport Assay Reagent Set (Gibco™)

The MDR1 Vesicular Transport Assay Reagent Set is a ready-to-use kit containing all reagents needed for vesicular transport assay except for the vesicles themselves. The reagent set can be used for vesicular transport assay using radioisotope-labeled or non-labeled compound. This kit has enough material to perform 100 vesicular transport assays.

Use The MDR1 Vesicular Transport Assay Reagent Set to:

• Investigate the transporter interactions of your drug candidates
• Assess potential for transporter-mediated drug-drug interactions
• Obtain high quality results with a large signal to noise ratios

Concept Behind ABC Transporter Vesicles
ABC transporter vesicles are easy-to-use, efficient reagents for early assessment of a drug candidate’s substrate and drug interaction potential. While ABC transporters typically mediate the export of substrates out of cells, transporters expressed on these inside-out vesicles import substrates into the vesicles. It is therefore possible to quantitatively evaluate transport activity for your compound by determining the amount incorporated into the vesicles.

Clear and Reliable Results
Prepared from Sf9 cells which have been engineered to over-express specific ABC transporters, these "inside-out" vesicles provide high levels of transporter activity with low background, giving you a clear signal if your compound is a substrate or inhibitor of a specific efflux transporter.

Kit Compatibility
Our MDR1 Vesicular Transport Assay Reagent Set is designed for use with MDR1 Vesicles. The 100 vesicular transport assays that can be performed with this kit will require two MDR1 Vesicle products.

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

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.

eBioscience™ BrdU Staining Kit for Flow Cytometry PerCP-eFluor™ 710 (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
PerCP-eFluor 710

Laser
Blue Laser

Emit
710 nm

Excite
488 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis