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Click-iT™ EdU Imaging Kit with Alexa Fluor™ 488, 594, and 647 Azides (Invitrogen™)

The Click-iT® EdU Imaging Kit is a trial version of our superior alternative to traditional proliferation assays optimized for fluorescence microscopy 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. This trial-size kit provides sufficient EdU to label up to 6–30 coverslips and enough Alexa Fluor® 488, 594, and 647 azide to detect new DNA synthesis on 2–10 coverslips with each fluorophore.
• 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 Imaging Kit 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 Imaging Kit 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.

This kit is optimized for fluorescence microscopy applications; visit the Click-iT® technology area of our website for kits designed for high-content imaging, fluorescence 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 EdU administration by feeding or injection methods.
The Click-iT® assay can be used with BrdU in dual pulse experiments by using the anti-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.

Click-iT™ AHA Alexa Fluor™ 488 Protein Synthesis HCS Assay (Invitrogen™)

The Click-iT® AHA Alexa Fluor® 488 Protein Synthesis HCS Assay provides a fast, sensitive, non-toxic and non-radioactive method for the detection of nascent protein synthesis utilizing fluorescence microscopy and high-content imaging. Click-iT® AHA is an amino acid analog of methionine containing an azido moiety. Similar to 35S-methionine, Click-iT® AHA is fed to cultured cells and incorporated into proteins during active protein synthesis. Detection of the incorporated amine acid utilizes a chemoselectiove ligation or “click" reaction between an azide and an alkyne, where the azido modified protein is detected with the green-fluorescent Alexa Fluor® 488 alkyne. The Click-iT® AHA Alexa Fluor® 488 Protein Synthesis HCS Assay has been successfully tested in A549 and U-2 OS cells with a variety of reagents that inhibit protein synthesis including cycloheximide, anisomycin and puromycin in both dose response and Min⁄Max format.

Click-iT™ EdU Microplate Assay (Invitrogen™)

The Click-iT® EdU Microplate Assay is a superior alternative to traditional cell proliferation assays that is optimized for microplate-based fluorescence analysis. This assay provides a simple and rapid workflow with reduced wash steps compared to traditional BrdU colorimetric or fluorescent proliferation assays. In this assay the modified thymidine analogue EdU is efficiently incorporated into newly synthesized DNA and fluorescently labeled with bright, photostable Amplex® UltraRed 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
• Sensitive—outstanding sensitivity and dynamic range
• Versatile—fluorescence or absorbance readouts through use of Amplex® UltraRed reagent

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.

Superior Proliferation Methodology
The Click-iT® EdU Microplate 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. The signal amplification steps include incubation with an anti-Oregon Green® antibody conjugated to horseradish peroxidase (HRP), which then reacts with Amplex® UltraRed substrate in a 1:1 stoichiometry and produces a bright red, fluorescent product (excitation/emission maxima ~568/585 nm).

With a high extinction coefficient, good quantum efficiency, and resistance to auto-oxidation, Amplex® UltraRed reagent delivers higher sensitivity and a broader assay range than other fluorogenic or colorimetric peroxidase substrates. Amplex® UltraRed reagent also offers versatility through detection by either fluorescence or absorbance measurement.

This kit also includes Amplex® UltraRed stop reagent that enables the peroxidase reaction to be terminated at a user-determined time point and stabilizes the signal for up to 24 hours. As a consequence, the Click-iT® EdU Microplate 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 Click-iT® EdU Microplate Assay is optimized for microplate-based fluorescence analysis; visit the Click-iT® technology area of our website for kits designed for fluorescence microscopy, high-content imaging, 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 anti-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.

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.

Arcturus™ Paradise™ PLUS 1.5 Round Kit, no solvents (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.

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.

Click-iT™ Plus EdU Pacific Blue™ Flow Cytometry Assay Kit (Invitrogen™)

The Click-iT® Plus EdU Pacific Blue™ Flow Cytometry Assay Kit provides a simplified, more robust assay for analyzing DNA replication in proliferating cells compared to traditional BrdU methods. Newly synthesized DNA is analyzed using the 405 nm laser of the flow cytometer. The Click-iT® Plus formulation is compatible with standard fluorophores, including R-PE and R-PE tandems, as well as fluorescent proteins.

• Multiplexable—compatible with R-PE (and tandems) and fluorescent proteins
• Accurate—superior results compared to BrdU assays
• Fast—results in as little as 60 minutes

View selection guide for all Click-iT™ EdU and Click-iT™ Plus EdU assays for flow cytometry.

Multiplexable
The Click-iT® Plus formulation provides increased multiplexibility compared to the original Click-iT® EdU Flow Cytometry assays. Click-iT® Plus EdU assays can be used in conjunction with R-PE and R-PE tandems, as well as fluorescent proteins such as GFP and mCherry, without loss of the accuracy or speed of the original Click-iT® EdU assay.

An Advanced Method Giving You Results Superior to BrdU
The most accurate method of proliferation analysis is direct measurement of DNA synthesis. Originally, this was performed through incorporation of radioactive nucleosides. This method was replaced by antibody-based detection of the nucleoside analog bromodeoxyuridine (BrdU). The Click-iT® Plus EdU Flow Cytometry assay is a novel alternative to the BrdU assay. EdU (5-ethynyl-2´-deoxyuridine) is a thymidine analog that is incorporated into DNA during active DNA synthesis. Detection is based on click chemistry, which is a copper-catalyzed covalent reaction between an azide and an alkyne. In this application, the alkyne is found in the ethynyl moiety of EdU, while the azide is coupled to the Alexa Fluor® dye. Standard flow cytometry methods are used for determining the percentage of S-phase cells in the population.

Mild Conditions Allow Use with Cell Cycle Dyes and Antibodies
The small size of the dye azide allows for efficient detection of the incorporated EdU using mild conditions, while standard aldehyde-based fixation and detergent permeabilization are sufficient for the Click-iT® Plus detection reagent to gain access to the DNA. This is in contrast to BrdU assays that require DNA denaturation (using HCl, heat, or digestion with DNase) to expose the BrdU so that it can be detected with an anti-BrdU antibody. Sample processing for the BrdU assay can result in signal alteration of the cell cycle distribution, as well as destruction of antigen recognition sites when using the HCl method. In contrast, the easy-to-use Click-iT® Plus EdU assay is compatible with cell cycle dyes. The Click-iT® Plus EdU assay can also be multiplexed with antibodies against surface and intracellular markers, as well as conjugates labeled with standard fluorophores including R-PE, R-PE tandems, and fluorescent proteins (GFP and mCherry).

Quick and Simple Protocol
The Click-iT® Plus EdU protocol is based on the aldehyde fixation and detergent permeabilization steps for immunohistochemical antibody labeling. However, EdU is compatible with other fixation/permeabilization agents including saponin and methanol. In just five steps you’ll be ready to analyze your cell proliferation data:
1. Treat cells with EdU.
2. Fix and permeabilize cells.
3. Detect S-phase cells with Click-iT® Plus detection cocktail for 30 min.
4. Wash once.
5. Analyze.
Results can be seen in as little as 60 minutes in some circumstances, but we recommend 90 minutes for all applications.

ABC Peroxidase Standard Staining Kit (Thermo Scientific™)

Thermo Scientific Pierce ABC Staining Kits include reagents for the avidin-biotin complex (ABC) technique, a highly sensitive method for immunohistochemical detection with biotinylated secondary antibodies.

Pierce ABC Staining Kits are highly sensitive, produce very low background staining and have rapid avidin-biotin interactions. Standard Kits contain only the two essential reagents (avidin and biotinylated HRP) can be used with nearly any species of biotinylated secondary antibody. The Ultra-Sensitive ABC Peroxidase Staining Kits are approximately five times more sensitive than the regular-senstivity kit, enabling more dilute primary antibodies to be used while producing equal staining intensity. Ultra-Sensitive Kits are offered as a Standard Kit and as Complete Kits, which contain biotinylated secondary antibodies and appropriate block serum for use with mouse or rabbit antibodies, respectively.

Related Products
Ultra-Sensitive ABC Peroxidase Standard Staining Kit
Ultra-Sensitive ABC Peroxidase Mouse IgG Staining Kit
Ultra-Sensitive ABC Peroxidase Rabbit IgG Staining Kit

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.

Arcturus™ Paradise™ PLUS qRT-PCR Kit, no solvents (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.

Arcturus™ Paradise™ PLUS 2 Round Kit (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.

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.

PrestoBlue™ Cell Viability Reagent (Invitrogen™)

Get high quality cell viability results and instant time savings with PrestoBlue® Cell Viability Reagent. PrestoBlue® is a ready to use cell permeable resazurin-based solution that functions as a cell viability indicator by using the reducing power of living cells to quantitatively measure the proliferation of cells. When added to cells, the PrestoBlue® reagent is modified by the reducing environment of the viable cell and turns red in color, becoming highly fluorescent. This color change can be detected using fluorescence or absorbance measurements.

The PrestoBlue® Cell Viability Reagent allows you to:

• Save time with an incubation step as short as 10 minutes.
• Obtain high quality results with a large dynamic range
• Experience convenience with the add and read homogenous format
• Monitor living cells and perform downstream functional assays


10 Minute Incubation
The PrestoBlue® Cell Viability Reagent is the only live-cell viability reagent on the market able to provide robust data typically in as little as 10 minutes following reagent addition. All other resazurin-based cell viability reagents on the market require a one to four hour incubation with the reagent prior to obtaining data.

High Quality Results
Based on the cell type and incubation time, PrestoBlue® Cell Viability Reagent can detect as few as 12 cells per well in a 384 well plate. After ten minutes of incubation the PrestoBlue® Reagent can linearly detect cells down to 98 cells⁄well with a Z’ value of greater than 0.5.

Homogenous Assay Format
The PrestoBlue® Cell Viability Reagent is an addition-only reagent. Without the cell lysis or solubilization steps required by other assays on the market, hands-on time is minimized.

Live Cell Assay
The PrestoBlue® Cell Viability Reagent does not require cell lysis. Cells may be incubated for up to 24 hours in the presence of PrestoBlue® Cell Viability Product. Simply remove the reagent from the cells and replace it with growth medium for further proliferation.

HCS LIVE/DEAD™ Green Kit (Invitrogen™)

The HCS LIVE⁄DEAD® Green Kit measures cytotoxicity by high content analysis with the Image-iT® DEAD Green™ viability stain – a non-fluorescent and cell impermeant compound that exhibits a strong fluorescent enhancement upon binding to DNA. Drugs and test compounds leading to serious cell injuries, including plasma membrane permeability, allow entry of the Image-iT® DEAD Green™ viability stain. This two-color fluorescence assay also employs the use of a cell permeant nuclear segmentation tool that stains both live and dead cells, reflecting the total cell number within the sample. For total cell staining, the kit includes both a blue-fluorescent Hoechst 33342 and a near infrared-fluorescent, HCS NuclearMask™ Deep Red reagent. Either of these segmentation tools may be used with Image-iT® DEAD Green™ viability stain for excellent discrimination of live and dead cells that survives fixation and permeabilization for combination with additional parameters of cytotoxicity or other biomarkers. The kit contains sufficient reagents for performing 2, 96-well plates.

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

Click-iT™ Plus EdU Cell Proliferation Kit for Imaging, Alexa Fluor™ 488 dye (Invitrogen™)

Click-iT Plus EdU Cell Proliferation imaging kits have been optimized for fluorescence microscopy applications and are a superior alternative to traditional proliferation assays. In this assay the modified thymidine analogue EdU (5-ethynyl-2'-deoxyuridine, a nucleoside analog of thymidine) is efficiently incorporated into newly synthesized DNA and fluorescently labeled with a bright, photostable Alexa Fluor® dye in a fast, highly-specific, mild click reaction.

Find more tools for image-based detection of proliferating cells >

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

The kit contains all of the components needed to label and detect the incorporated EdU as well as perform cell cycle analysis on samples from adherent cells. For cell cycle analysis, the kit includes a blue fluorescent Hoechst 33342 dye. The kit contains sufficient reagents for labeling 50 18x18 coverslips using 500 µL of reaction buffer per test.

Avoid the Harsh Treatments Associated with BrdU Method
Measuring changes to cell proliferation is a fundamental method for assessing cell health, determining genotoxicity, and evaluating anti-cancer drugs. The most accurate method of doing this is by directly measuring DNA synthesis. The traditional method utilizes the nucleoside analog BrdU (5-bromo-2'-deoxyuridine, a nucleoside analog of thymidine), which is incorporated into newly transcribed DNA. After incorporation the samples are treated with harsh methods (HCl, heat, or enzymes) to denature the DNA and expose the BrdU molecules to detection by anti-BrdU antibodies. However, the BrdU method to measure cell proliferation is time consuming and difficult to perform consistently. The harsh treatments necessary for this method can adversely affect sample integrity, cell morphology, image quality, and the ability to multiplex.

The Click-iT® Plus EdU assay measures the rate of new DNA synthesis based on incorporation of the nucleoside analog EdU into DNA. Detection is achieved through a catalyzed "click" reaction that is completed typically within 30 minutes. The click reaction uses bioorthogonal (biologically unique) moieties to fluorescently label proliferating cells, producing low backgrounds and high detection sensitivities. Because of the mild reaction conditions the Click-iT® Plus assays can accurately determine cell proliferation while preserving cell morphology, DNA integrity, antigen binding sites, and the fluorescent signal from GFP. Preservation of DNA integrity allows for DNA staining, including staining with dyes used for cell cycle analysis.

LIVE/DEAD™ Cell Imaging Kit (488/570) (Invitrogen™)

The LIVE/DEAD® Cell Imaging Kit is a sensitive two-color fluorescence cell viability assay optimized for FITC and Texas Red® filters. Quick and easy to use, the kit allows discrimination between live and dead cells with two probes that measure recognized parameters of cytotoxicity and cell viability—intracellular esterase activity and plasma membrane integrity.

The LIVE/DEAD® Cell Imaging Kit *488/570* offers:

Fast, simple determination of live and dead cells

• Accuracy with convenience
• Sensitive probes ideal for FITC and Texas Red filters


Dual Probe-Based Assay for Imaging Platforms
Just like our popular LIVE/DEAD® Viability/Cytotoxicity assay, the new LIVE/DEAD® Cell Imaging Kit is based on a cell-permeable dye for staining of live cells and a cell-impermeable dye for staining of dead and dying cells, which are characterized by compromised cell membranes. To adapt this important assay for imaging platforms, the LIVE/DEAD® Cell Imaging Kit components were optimized for the common green and red imaging filters used with FITC and Texas Red. The live cell component produces an intense, uniform green fluorescence in live cells (ex/em 488 nm/515 nm). The dead cell component produces a predominantly nuclear red fluorescence (ex/em 570nm/602 nm) in cells with compromised cell membranes, a strong indicator of cell death and cytotoxicity (Fig 1).

Quick, Exact Determination of Viability and Cytotoxicity
The LIVE/DEAD® Cell Imaging kit provides very sensitive detection of key measures of cell health: viability, live/dead ratio, and cytotoxicity. The components of the LIVE/DEAD® Cell Imaging kit are configured for ease of use with minimal dilutions.

Assay Principle
With the LIVE/DEAD® Cell Imaging Kit, live cells are distinguished by the presence of ubiquitous intracellular esterase activity as determined by the enzymatic conversion of the virtually non-fluorescent cell-permeant calcein AM to the intensely fluorescent calcein, which is well-retained within live cells. The red component of the LIVE/DEAD® Cell Imaging Kit is cell-impermeant and therefore only enters cells with damaged membranes. In dying and dead cells a bright red fluorescence is generated upon binding to DNA. Background fluorescence levels are inherently low with this assay technique because the dyes are virtually non-fluorescent before interacting with cells. The fluorophores in the LIVE/DEAD® Cell Imaging Kit were selected for their brightness, spectral properties (FITC and Texas Red filters), and ease of use. Packaged for workflow convenience, they allow for effortless and quick determination of the fractions of live and dead cells in a population.

See other Molecular Probes imaging tools and reagents