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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.

Click-iT™ Plus TUNEL Assay for In Situ Apoptosis Detection, Alexa Fluor™ 488 dye (Invitrogen™)

The Click-iT® Plus TUNEL assay detects apoptotic cells in tissue and cultured cell samples through the use of a small, highly specific labeling moiety and a bright fluorescent dye. After incorporation of the labeling moiety into DNA fragments, detection is achieved through a catalyzed “click” reaction using conditions mild enough to preserve the fluorescent signal from GFP or RFP.

More tools for apoptosis detection and measurement >

• Optimized for the detection of apoptotic cells in either tissue or cell samples
• Multiplex enabled—optimized to work with fluorescent dyes or proteins such as GFP and RFP
• Improved TUNEL assay—better label incorporation due to small reactive moiety
• Bright apoptotic signal—uses Alexa Fluor® dyes, resulting in a stable, non-photobleaching fluorescent signal
• Flexibility—the assay can be configured to test from 1 to 50 samples at a time

Fragmentation of cellular DNA is a hallmark of apoptosis. The TUNEL assay is the most widely used method to detect fragmented DNA in apoptotic cells in tissue samples. The TUNEL assay begins with incorporation of modified dUTP at the 3’-OH end of the fragmented DNA. The dUTP modification is often the addition of a fluorophore. And due to the size of the fluorophore, the modified dUTP can display lower than expected incorporation rates. These lower incorporation rates can affect the sensitivity of the TUNEL assay. Additionally, the fluorophores used in currently available TUNEL assays suffer from photobleaching and fluorescent spectral overlap issues, both of which reduce sensitivity and ability to multiplex the assay.

The Click-iT® Plus TUNEL assay was developed to address these issues. The assay uses dUTP modified with an alkyne group (a small bio-orthogonal functional group) that allows the nucleotide to be more readily incorporated. After incorporation, a highly specific click reaction between the alkyne group and an Alexa Fluor® picolyl azide fluorescent dye, and subsequent detection of that dye, results in a sensitive and specific assay for the detection of apoptotic cells. Because of the gentle reaction conditions, the Click-iT® Plus TUNEL assay enables multiplexing with fluorescent proteins or dyes.

The Click-iT® Plus TUNEL assay has been validated with several different formalin-fixed paraffin-embedded tissue types. In all cases the ability to multiplex with fluorescent proteins and dyes was preserved. Additionally, the ability to stain actin using fluorescent-labeled phalloidin was also preserved.

The Click-iT® Plus TUNEL assay contains all of the reagents needed to detect apoptotic cells from either tissue or cell samples. The reagents supplied in this kit can be used to test 50 samples and can be configured to test from 1 to 50 samples at a time.

Arcturus™ Paradise™ PLUS 2 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.

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.

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

eBioscience™ Annexin V Apoptosis Detection Kit PE (Invitrogen™)

Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric distribution across the phospholipid bilayer and is translocated to the extracellular membrane leaflet marking cells as targets of phagocytosis. Once on the outer surface of the membrane, PS can be detected by fluorescently labeled Annexin V in a calcium-dependent manner.

In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD, or Fixable Viability Dyes such as eFluor™ 660 or eFluor™ 780. These cells will stain with Annexin V but not a viability dye, thus distinguishing cells in early apoptosis. However, in late stage apoptosis, the cell membrane loses integrity thereby allowing Annexin V to also access PS in the interior of the cell. A viability dye can be used to resolve these late-stage apoptotic and necrotic cells (Annexin V, viability dye-positive) from the early-stage apoptotic cells (Annexin V positive, viability dye-negative).

Note: Fixable Viability Dye eFluor™ 450 is not recommended for use with Annexin V Apoptosis Detection Kits.

Reported Application
Flow Cytometric Analysis

ATP Determination Kit, 200-1,000 assays (Invitrogen™)

Molecular Probes® ATP Determination Kit offers a convenient bioluminescence assay for quantitative determination of ATP with recombinant firefly luciferase and its substrate D-luciferin. The assay is based on luciferase’s absolute requirement for ATP in producing light (emission maximum ~560 nm at pH 7.8). This assay is extremely sensitive; most luminometers can detect as little as 0.1 picomole of pre-existing ATP, or ATP as it is being formed in kinetic systems. This sensitivity has led to numerous applications for detecting ATP production in various enzymatic reactions, including ATPase and NADPH oxidase, as well as for detecting low-level bacterial contamination in samples such as blood, milk, urine, soil and sludge. The luciferin–luciferase bioluminescence assay has also been used successfully to study the effects of antibiotics on bacterial populations, to determine cell proliferation and cytotoxicity in both bacterial and mammalian cells, and to distinguish cytostatic versus cytocidal potential of anticancer drugs on malignant cell growth.

ATP Determination Kit Specifications:
• Luminescence is monitored at ~560 nm using a luminometer
• Can detect as little as 0.1 picomole of pre-existing ATP, or ATP as it is being formed in kinetic systems
• Luciferase and luciferin are packaged separately, which enables researchers to optimize conditions for their samples and instruments
• Includes sufficient reagents to perform 200 ATP assays using 500 µL sample volumes or 1,000 ATP assays using 100 µL sample volumes


Find More Viability and Cytotoxicity Assays and Probes for Detecting Enzymes That Metabolize Phosphates
Review Viability and Cytotoxicity Assay Kits for Diverse Cell Types—Section 15.3 and Detecting Enzymes That Metabolize Phosphates and Polyphosphates—Section 10.3 in the Molecular Probes® Handbook for more information on these products.

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

Click-iT™ Plus TUNEL Assay for In Situ Apoptosis Detection, Alexa Fluor™ 594 dye (Invitrogen™)

The Click-iT® Plus TUNEL assay detects apoptotic cells in tissue and cultured cell samples through the use of a small, highly specific labeling moiety and a bright fluorescent dye. After incorporation of the labeling moiety into DNA fragments, detection is achieved through a catalyzed “click” reaction using conditions mild enough to preserve the fluorescent signal from GFP or RFP.

More tools for apoptosis detection and measurement >

• Optimized for the detection of apoptotic cells in either tissue or cell samples
• Multiplex enabled—optimized to work with fluorescent dyes or proteins such as GFP and RFP
• Improved TUNEL assay—better label incorporation due to small reactive moiety
• Bright apoptotic signal—uses Alexa Fluor® dyes, resulting in a stable, non-photobleaching fluorescent signal
• Flexibility—the assay can be configured to test from 1 to 50 samples at a time

Fragmentation of cellular DNA is a hallmark of apoptosis. The TUNEL assay is the most widely used method to detect fragmented DNA in apoptotic cells in tissue samples. The TUNEL assay begins with incorporation of modified dUTP at the 3’-OH end of the fragmented DNA. The dUTP modification is often the addition of a fluorophore. And due to the size of the fluorophore, the modified dUTP can display lower than expected incorporation rates. These lower incorporation rates can affect the sensitivity of the TUNEL assay. Additionally, the fluorophores used in currently available TUNEL assays suffer from photobleaching and fluorescent spectral overlap issues, both of which reduce sensitivity and ability to multiplex the assay.

The Click-iT® Plus TUNEL assay was developed to address these issues. The assay uses dUTP modified with an alkyne group (a small bio-orthogonal functional group) that allows the nucleotide to be more readily incorporated. After incorporation, a highly specific click reaction between the alkyne group and an Alexa Fluor® picolyl azide fluorescent dye, and subsequent detection of that dye, results in a sensitive and specific assay for the detection of apoptotic cells. Because of the gentle reaction conditions, the Click-iT® Plus TUNEL assay enables multiplexing with fluorescent proteins or dyes.

The Click-iT® Plus TUNEL assay has been validated with several different formalin-fixed paraffin-embedded tissue types. In all cases the ability to multiplex with fluorescent proteins and dyes was preserved. Additionally, the ability to stain actin using fluorescent-labeled phalloidin was also preserved.

The Click-iT® Plus TUNEL assay contains all of the reagents needed to detect apoptotic cells from either tissue or cell samples. The reagents supplied in this kit can be used to test 50 samples and can be configured to test from 1 to 50 samples at a time.

Arcturus™ Paradise™ PLUS QC Kit (Applied Biosystems™)

Formalin-fixed paraffin-embedded (FFPE) samples introduce unique challenges for gene expression profiling and gene expression quantitation, including chemical modification and fragmentation of RNA molecules. Archival FFPE samples present additional challenges due to increased RNA degradation over time. As a result, not all FFPE samples contain high quality RNA.

The ARCTURUS® Paradise® PLUS Sample Quality Assessment Kit offers a simple solution for evaluating the quality of RNA from Formalin-Fixed Paraffin-Embedded (FFPE) tissues before proceeding with laser microdissection and downstream molecular analysis. The reagents provided in the kit are optimized to enable efficient extraction, isolation, and reverse transcription of total RNA from FFPE tissue scrapes. The RNA quantity and quality is then measured using quantitative real-time PCR (Figure 1).

The RNA quantity and quality are assessed by the presence of intact RNA for a specific message. Two sets of β-actin gene-specific primers designed to amplify short segments of DNA within the β-actin target sequence are used to estimate the amount of RNA in a given sample in relation to β-actin content using the real-time PCR technique. The RNA quantity derived from the 3' primer set is used to quantitate the RNA in the FFPE tissue scrape. The ratio of the RNA yield obtained from both sets of PCR primers is the 3'⁄5' ratio and is used as an indication of RNA quality.

Key product features:
• Secure Downstream Success with the Paradise® QC Kit
• Benefit from the Only Complete System for Formalin-fixed 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 FFPE 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 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.

Click-iT™ Plus TUNEL Assay for In Situ Apoptosis Detection, Alexa Fluor™ 647 dye (Invitrogen™)

The Click-iT® Plus TUNEL assay detects apoptotic cells in tissue and cultured cell samples through the use of a small, highly specific labeling moiety and a bright fluorescent dye. After incorporation of the labeling moiety into DNA fragments, detection is achieved through a catalyzed “click” reaction using conditions mild enough to preserve the fluorescent signal from GFP or RFP.

More tools for apoptosis detection and measurement >

• Optimized for the detection of apoptotic cells in either tissue or cell samples
• Multiplex enabled—optimized to work with fluorescent dyes or proteins such as GFP and RFP
• Improved TUNEL assay—better label incorporation due to small reactive moiety
• Bright apoptotic signal—uses Alexa Fluor® dyes, resulting in a stable, non-photobleaching fluorescent signal
• Flexibility—the assay can be configured to test from 1 to 50 samples at a time

Fragmentation of cellular DNA is a hallmark of apoptosis. The TUNEL assay is the most widely used method to detect fragmented DNA in apoptotic cells in tissue samples. The TUNEL assay begins with incorporation of modified dUTP at the 3’-OH end of the fragmented DNA. The dUTP modification is often the addition of a fluorophore. And due to the size of the fluorophore, the modified dUTP can display lower than expected incorporation rates. These lower incorporation rates can affect the sensitivity of the TUNEL assay. Additionally, the fluorophores used in currently available TUNEL assays suffer from photobleaching and fluorescent spectral overlap issues, both of which reduce sensitivity and ability to multiplex the assay.

The Click-iT® Plus TUNEL assay was developed to address these issues. The assay uses dUTP modified with an alkyne group (a small bio-orthogonal functional group) that allows the nucleotide to be more readily incorporated. After incorporation, a highly specific click reaction between the alkyne group and an Alexa Fluor® picolyl azide fluorescent dye, and subsequent detection of that dye, results in a sensitive and specific assay for the detection of apoptotic cells. Because of the gentle reaction conditions, the Click-iT® Plus TUNEL assay enables multiplexing with fluorescent proteins or dyes.

The Click-iT® Plus TUNEL assay has been validated with several different formalin-fixed paraffin-embedded tissue types. In all cases the ability to multiplex with fluorescent proteins and dyes was preserved. Additionally, the ability to stain actin using fluorescent-labeled phalloidin was also preserved.

The Click-iT® Plus TUNEL assay contains all of the reagents needed to detect apoptotic cells from either tissue or cell samples. The reagents supplied in this kit can be used to test 50 samples and can be configured to test from 1 to 50 samples at a time.

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.

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

The Click-iT® EdU Pacific Blue™ Flow Cytometry Assay Kit provides a simplified, more robust assay for analyzing DNA replication in proliferating cells as compared to traditional BrdU methods. Newly synthesized DNA is analyzed using the 405 nm laser of the flow cytometer.

Accurate—superior results compared to BrdU assays
Fast—results in as little as 90 minutes
Economical—more assays per kit

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

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, i.e., 3H-thymidine. This method was replaced by antibody-based detection of the nucleoside analog bromodeoxyuridine (BrdU). The Click-iT® EdU Flow Cytometry Assay Kits are novel alternatives to the BrdU assay. EdU (5-ethynyl-2´-deoxyuridine) is a thymidine analog which is incorporated into DNA during active DNA synthesis. Detection is based on click chemistry: 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 Alexa Fluor ® 488, Alexa Fluor® 647, or Pacific Blue™ dyes. Standard flow cytometry methods are used for determining the percentage of S-phase cells in the population (Fig 1).

Mild Conditions Allow Use with Cell Cycle Dyes and Antibodies
The advantages of Click-iT® EdU labeling are readily evident while performing the assay (Fig 2). 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® 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 may 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 EdU cell proliferation kit is compatible with cell cycle dyes. This EdU assay kit can also be multiplexed with antibodies against surface and intracellular markers.

Quick and Simple Protocol
The Click-iT® EdU protocol is based on the aldehyde fixation and detergent permeabilization steps for immunohistochemical antibody labeling, but 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:

• Treat cells with EdUM
• Fix and permeabilize cells
• Detect S-phase cells with Click-iT® detection cocktail for 30 min
• Wash once
• Analyze

Get Accurate Results Economically
By increasing the number of assays per kit, the Click-IT® EdU Pacific Blue™ Flow Cytometry Assay Kit is less expensive than the traditional BrdU assays making them ideal for large experiments.

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

eBioscience™ Annexin V Apoptosis Detection Set PE-Cyanine7 (Invitrogen™)

Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS) in all mammalian species. Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric distribution across the phospholipid bilayer and is translocated to the extracellular membrane leaflet marking cells as targets of phagocytosis. Once on the outer surface of the membrane, PS can be detected by fluorescently labeled Annexin V in a calcium-dependent manner.

In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD, or Fixable Viability Dyes such as eFluor™ 660 or eFluor™ 780. These cells will stain with Annexin V but not a viability dye, thus distinguishing cells in early apoptosis. However, in late stage apoptosis, the cell membrane loses integrity thereby allowing Annexin V to also access PS in the interior of the cell. A viability dye can be used to resolve these late-stage apoptotic and necrotic cells (Annexin V, viability dye-positive) from the early-stage apoptotic cells (Annexin V positive, viability dye-negative).

Due to the emission spectrum of PE-Cyanine7, the Annexin V Apoptosis Detection Set PE-Cyanine7 is not compatible with propidium iodide and 7-AAD. It is recommended to substitute a Fixable Viability Dye such as eFluor™ 660 or eFluor™ 780 in their place.

Not included:
Fixable Viability Dye eFluor™ 660 (cat. 65-0864)
Fixable Viability Dye eFluor™ 780 (cat. 65-0865)

Note: Fixable Viability Dye eFluor™ 450 is not recommended for use with Annexin V Apoptosis Detection Kits.

Reactivity/Species
Human, Mouse, Rat

Reported Application
Flow Cytometric Analysis

Click-iT™ TUNEL Colorimetric IHC Detection Kit (Invitrogen™)

The Click-iT™ TUNEL Colorimetric IHC Detection Kit is used to identify apoptotic cells in tissue and cultured cell samples through the use of a small, highly specific labeling moiety and streptavidin-peroxidase conjugation. After incorporation of the labeling moiety into sites of DNA fragmentation, detection is achieved through a catalyzed "click" reaction that adds a biotin group at these sites. The subsequent addition of a streptavidin-peroxidase and peroxidase substrate results in a dark brown signal that can be detected with light microscopy and stored for future analysis.

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

• Optimized for the detection of apoptotic cells in either tissue or cell samples
• Improved colorimetric TUNEL assay—better label incorporation due to small reactive moiety
• Increased sensitivity—specific label incorporation results in lower background and brighter signal
• Content-rich results—cell morphology, cellular environment, and apoptotic signal are clearly visible
• Flexibility—the assay can be configured for 50 independent TUNEL apoptosis tests

The later stages of apoptosis are characterized by changes in nuclear morphology, chromatin condensation, nuclear envelope degradation, and DNA fragmentation. Terminal deoxynucleotidyl transferase-dUTP nick end labeling (TUNEL) assays are based on the incorporation of modified dUTPs by terminal deoxynucleotidyl transferase (TdT) at the 3’-OH ends of fragmented DNA. Colorimetric TUNEL assays utilize dUTPs conjugated with a biotin moiety, followed by the addition of a streptavidin-peroxidase conjugate and peroxidase substrate, resulting in a dark brown apoptotic signal. However, colorimetric TUNEL assays are prone to high background, which reduces the sensitivity and specificity of the signal.

The Click-iT TUNEL Colorimetric IHC Detection Kit was developed to address these issues by utilizing a dUTP modified with an alkyne group (a small bio-orthogonal functional group) that enables the nucleotide to be more readily incorporated by TdT. After incorporation, a highly specific click reaction covalently links biotin azide and the alkyne group. Streptavidin-peroxidase (horseradish peroxidase) followed by peroxidase substrate (DAB) are added, allowing colorimetric detection of apoptotic cells. The high degree of labeling specificity inherent in the click technology results in low background and improved detection of apoptotic cells.

The Click-iT TUNEL Colorimetric IHC Detection Kit has been optimized and contains all of the reagents needed for detection of apoptotic cells from either tissue or cell samples. The reagents supplied in this kit can be used to test 50 samples and configured to test from one to 50 samples at a time.

Tali™ Apoptosis Kit - Annexin V Alexa Fluor™ 488 & Propidium Iodide (Invitrogen™)

The Tali® Apoptosis Kit enables identification of apoptotic cells and discrimination of apoptotic from necrotic and live cells in a population. The kit stains apoptotic cells with green Annexin V – Alexa Fluor® 488, stains necrotic cells with both red propidium iodide and green Annexin V – Alexa Fluor® 488, and does not stain live cells.

The Tali®Apoptosis Kit contains:

Annexin V – Alexa Fluor® 488 conjugate

• Annexin Binding solution
• Propidium Iodide (PI) solution

Apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. Inappropriately regulated apoptosis is implicated in disease states, such as Alzheimer’s disease and cancer. Apoptosis is distinguished from necrosis by characteristic morphological and biochemical changes, including compaction and fragmentation of the nuclear chromatin, shrinkage of the cytoplasm, and loss of membrane asymmetry.

Easily Discriminate Apoptotic from Necrotic Cells with Two-color Staining
In normal live cells, phosphatidyl serine (PS) is located on the cytoplasmic surface of the cell membrane. In apoptotic cells, however, PS is translocated from the inner to the outer surface of the plasma membrane, exposing it to the extracellular environment. The human anticoagulant Annexin V displays a high affinity for PS. Annexin V labeled with a fluorophore such as Alexa Fluor® 488 can be used to identify apoptotic cells by binding to the exposed PS.

The Tali® Apoptosis Kit includes a solution of propidium Iodide (PI). PI is a cell-impermeant fluorogenic DNA-binding dye used for years to identify necrotic cells. PI is impermeant to live cells, but easily enters dead cells where it binds to nucleic acids and becomes fluorescent.

After staining a cell population using the Tali® Apoptosis Assay Kit with Annexin V-Alexa Fluor® 488 and Propidium Iodide, apoptotic cells display green fluorescence, dead cells display red and green fluorescence and live cells show little or no fluorescence.

BSEP Vesicular Transport Assay Reagent Set (Gibco™)

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

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