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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.

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

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

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

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

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

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

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

Click-iT™ EdU Cell Proliferation Kit for Imaging, Alexa Fluor™ 647 dye (Invitrogen™)

The Click-iT EdU Cell Proliferation Kit for Imaging is a superior alternative to traditional proliferation assays that is 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.

• 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 Cell Proliferation Kit for Imaging 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 >
Find more tools for image-based detection of proliferating cells >

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.

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

Vybrant™ Cell Adhesion Assay Kit (Invitrogen™)

The Vybrant® Cell Adhesion Assay Kit is a fast and sensitive assay for measuring cell-cell or cell-surface adhesion for a variety of cell types. In this assay, cells are labeled with calcein AM and allowed to adhere. After removal of nonadherent cells, calcein fluorescence is used to calculate the number of adherent cells.

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.

Click-iT™ Plus EdU Cell Proliferation Kit for Imaging, Alexa Fluor™ 555 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.

eBioscience™ Annexin V Apoptosis Detection Kit eFluor™ 450 (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).

Reported Application
Flow Cytometric Analysis

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

The LIVE/DEAD™ Fixable Violet 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 Violet 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 Violet 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 Violet 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 violet-fluorescent reactive dye has an excitation maximum of ~416 nm and is excited well with the 405 nm violet laser, and it has an emission maxima of ~451 nm, so it can be collected in the first 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 2 Round Kit, Cy3 labeling (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.

LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation (Invitrogen™)

The LIVE/DEAD™ Fixable Near-IR 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 red laser flow cytometer.

• Stable—dyes are freeze dried in separate vials to maintain stability

• Robust—staining pattern is the same before and after fixation

• Bright signal—allows for easy distinction between live/dead cells in single channel

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

Stable
Unlike products that are sold in solution, the LIVE/DEAD™ Fixable Near-IR 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 Near-IR Stain is an amine reactive dye that binds covalently to intracellular and extracellular amines, and the staining pattern is preserved following formaldehyde fixation.

Optimal brightness
The LIVE/DEAD™ Fixable Near-IR stain was selected based on its fluorescent properties to provide a bright signal when excited with a red laser. The near-IR fluorescent reactive dye has an excitation maximum of ~633 nm making it ideal for use with the red or HeNe laser with an emission of ~750 nm. Since live and dead cells can be discriminated using a single dye and a single channel of a flow cytometer, it is an ideal choice for multi-color experiments.

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.

SuperPicture™ Polymer Detection Kit, HRP, broad spectrum

The SuperPicture™ Polymer Detection kit features our proprietary and enhanced HRP polymer technology to provide excellent sensitivity, high specificity, and a shorter
protocol than conventional immunohistochemical staining methods.

Our new polymer conjugation methods have resulted in a versatile polymer capable of intense nuclear, cytoplasmic, and membrane antigen staining. The polymer conjugate contains multiple HRP molecules to provide excellent sensitivity. The polymer staining method is faster and easier than a conventional LAB-SA protocol because it uses an HRP polymer conjugated to the second antibody and eliminates one incubation step. In addition, SuperPicture™ Polymer kits do not contain biotin nor streptavidin, so potential background due to endogenous biotin activity is completely avoided. Another advantage of SuperPicture™ Polymer detection system is the shorter polymer incubation time; the this system only requires 10 minutes, while leading competitors’ polymers require 30 minutes.

Reactivity: Mouse, Rabbit, Rat, G. Pig

Applications: Immunohistochemistry (FFPE)

BacLight™ RedoxSensor™ Green Vitality Kit (Invitrogen™)

The RedoxSensor™ Green reagent included in the BacLight™ RedoxSensor™ Green Vitality Kit is an indicator of bacterial reductase activity. This reductase activity is, in turn, a reliable marker for changes in electron transport chain function and for changes in vitality that occurs following antibiotic treatment. RedoxSensor™ Green reagent penetrates both gram-positive and gram-negative bacteria. Following reduction, the RedoxSensor™ Green reagent will produce a stable green-fluorescent signal in 10 minutes that is compatible with formaldehyde fixation techniques.

View additional information about all microbiology assays for flow cytometry.

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

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

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

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

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

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

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

Tali™ Viability Kit - Dead Cell Red (Invitrogen™)

The Tali® Viability Kit - Dead Cell Red is a ready-to-use solution of propidium iodide (PI) that has been validated for use with the Tali® Image-Based Cytometer. PI is a live cell-impermeant fluorogenic DNA-binding dye that has been extensively used to identify necrotic cells.

Key Features of Tali® Viability Kit - Dead Cell Red:

• Impermeant to live cells
• Easily enters and binds to nucleic acids in necrotic cells

After staining a cell population using Dead Cell Red™, dead cells display red fluorescence upon excitation. Tali® Viability Kit - Dead Cell Red can be used to identify dead cells in a population of unstained cells, or in a population of GFP-expressing cells.