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eBioscience™ Annexin V Apoptosis Detection Kit APC (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

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)

LIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation (Invitrogen™)

The LIVE/DEAD™ Fixable Green 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 blue 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 Green 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 Green 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 Green stain was selected based on its fluorescent properties to provide a bright signal when excited with a blue 488 nm laser. The green-fluorescent reactive dye has an excitation maximum of ~495 nm making it ideal for use with the blue laser and an emission of ~520 nm allowing collection in the FITC channel. 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.

eBioscience™ Annexin V Apoptosis Detection Kit PerCP-eFluor™ 710 (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 PerCP-eFluor™ 710, the Annexin V Apoptosis Detection Kit PerCP-eFluor™ 710 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.

Reported Application
Flow Cytometric Analysis

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

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

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

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

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

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

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

Integrated Systems for Microgenomics
The Paradise® PLUS Reagent System is validated as part of ARCTURUS®' complete Systems for Microgenomics®, an integrated solution for utilizing small quantities of RNA for gene expression analysis, and features the ARCTURUSXT Laser Capture Microdissection (LCM) Instrument to procure pure cell populations. ARCTURUS®' Systems for Microgenomics permit accurate and sensitive microarray assays that reveal molecular otherwise obscured in whole tissue samples or cell mixtures.

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

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.

LC3B Antibody Kit for Autophagy (Invitrogen™)

The LC3 Antibody Kit for Autophagy includes a rabbit polyclonal antibody against LC3B that has been validated for use in fluorescence microscopy and high content imaging and analysis. Chloroquine diphosphate is also included - following treatment with this compound, normal autophagic flux is disrupted and autophagosomes accumulate as a result of the disruption in normal autophagic flux.

CellEvent™ Caspase-3/7 Green ReadyProbes™ Reagent (Invitrogen™)

CellEvent® Caspase-3/7 Green ReadyProbes® Reagent is a fluorogenic, no-wash indicator of activated caspase-3/7 for live- and fixed-cell applications. Activation of caspase-3 is an early indicator of apoptosis and CellEvent® Caspase-3/7 Green reagent allows rapid and sensitive detection of cells destined for cell death.

• Simple and fast no-wash protocol helps preserve delicate apoptotic cells
• Compatible with both live-cell fluorescence imaging and formaldehyde-based fixation methods
• Ready-to-use liquid formulation in convenient dropper bottle—no need to dilute, weigh, or pipette
• Stability at room temperature—keep handy at your scope or cell culture area.

See other ReadyProbes® reagents for cell staining
Learn more about other assays for apoptosis

Cell imaging applications
CellEvent® Caspase-3/7 Green reagent is a four amino acid peptide (DEVD) conjugated to a nucleic acid-binding dye that is non-fluorescent when not bound to DNA. The CellEvent® Caspase-3/7 Green reagent is intrinsically non-fluorescent, as the DEVD peptide inhibits binding of the dye to DNA. Upon activation of caspase-3/7 in apoptotic cells, the DEVD peptide is cleaved and the free dye can bind DNA, generating a bright green fluorescence. The fluorescence emission of the dye when bound to DNA is 530 nm and can be observed using a standard FITC filter set.

Suggestions for use
• In most cases, 2 drops/ml and an incubation time of 30 minutes is sufficient for bright nuclear staining of apoptotic cells; however, optimization may be needed for some cell types, conditions, and applications. In such cases, simply add more or fewer drops until the optimal staining intensity is obtained.
• If desired, combine CellEvent™ Caspase-3/7 Green ReadyProbes® reagent with a red or far-red nuclear stain for a total cell count
• CellEvent™ Caspase-3/7 Green dye is excited with a maximum at 502 nm and has an emission maximum at 530 nm. It is detected through standard GFP and FITC filters.

Arcturus™ Paradise™ PLUS 1.5 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.

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.

eBioscience™ BrdU Kit for IHC/ICC Immunofluorescence eFluor™ 660 (Invitrogen™)

This BrdU Kit for immunohistochemistry (IHC)/immunocytochemistry (ICC) with immunofluorescence detection contains the necessary reagents and buffers for identifying and examining proliferating cells by immunohistochemical or immunocytochemical analysis. Cells are labeled in vitro or in vivo with 5-bromo-2’-deoxyuridine (BrdU), a synthetic analog of thymidine, which is incorporated into DNA in place of thymidine during the S-phase of the cell cycle. Following fixation and antigen retrieval steps, cells or tissue sections are stained for BrdU incorporation and visualized using a streptavidin-conjugated fluorophore. This kit has been optimized for IHC with both frozen and formalin-fixed paraffin embedded BrdU-labeled mouse intestine and ICC of BrdU-pulsed HeLa cells grown on culture slides.

Conjugate
eFluor 660

Laser
Red Laser

Emit
668 nm

Excite
633 - 647 nm

Reported Application
Immunohistochemical Staining of Formalin-Fixed Paraffin Embedded Tissue Sections, Immunohistochemical Staining of Frozen Tissue Sections, Immunocytochemistry, Microscopy

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

CellTrace™ Yellow Cell Proliferation Kit is used for in vitro and in vivo labeling of cells to trace multiple generations using dye dilution by flow cytometry. This reagent is specifically designed for excitation by either a 532-nm or 561-nm laser, and it can be easily multiplexed with reagents and antibodies excited by other commonly used lasers.

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

Additional information is available about other CellTrace™ Reagents to meet your multiplexing needs.

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

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

Easy multiplexing with other fluorophores
The yellow excitation at 546 nm and emission at 579 nm of CellTrace Yellow dye make it ideal for multiplexing due to the limited spectral overlap with other common dyes (e.g., Alexa Fluor™488 and FITC) and green fluorescent protein (GFP) .

Use the SpectraViewer to help you plan your experiments.

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

Related links
General information about Thermo Fisher Scientific flow cytometry products and support is also available.

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.

FungaLight™ Yeast CFDA, AM/Propidium Iodide Vitality Kit (Invitrogen™)

The FungaLight™ Yeast CFDA, AM / Propidium Iodide Vitality Kit combines a cell permeable esterase substrate with a membrane integrity indicator to evaluate the vitality of yeast cells by flow cytometry or microscopy. The acetoxymethyl ester (AM) of the esterase substrate 5-carboxyfluorescein diacetate (CFDA) allows this reagent to permeant cell membranes Once inside the cell, the lipophilic blocking and diacetate groups are cleaved by nonspecific esterases, resulting in a fluorescent, charged form that leaks out of cells very slowly. In contrast, the membrane integrity indicator, propidium iodide penetrates yeast with damaged membranes. With an appropriate mixture of the CFDA, AM and propidium iodide stains, esterase-active yeast with intact cell membranes stain fluorescent green, whereas yeast with damaged membranes stain fluorescent red.

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

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

With the CyQUANT® Cell Proliferation Assay Kit, you can:

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

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

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

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

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

pHrodo™ Red Phagocytosis Particle Labeling Kit for Flow Cytometry (Invitrogen™)

• Specifically detect phagocytosis and endocytosis with pH-sensitive fluorogenic dye - discriminate endcytosed from adherent and extracellular particles
• Reduced signal variability and improved timing in sensitive experiments - no need for wash steps or quencher dye
• Bright red fluorescence - conveniently multiplex with green dyes such as GFP, Fluo-4, or calcein

View a selection guide for all pHrodo Indicators used in imaging, flow cytometry and microplate assays.

The pHrodo™ Red dye gives faster and more accurate results than any other phagocytosis assay

The new Molecular Probes™ proprietary pH-sensitive rhodamine-based pHrodo™ Red dye is non-fluorescent at neutral pH, but turns bright red upon acidification. Because it is both fluorogenic and pH-sensitive, the pHrodo™ Red dye can be used as a specific sensor of phagocytic events; acidification of the phagosome following phagocytosis is marked by red fluorescence. It is therefore an ideal tool with which to study phagocytosis and its regulation by drugs and/or environmental factors.

Rapid and reproducible

Wash steps and quencher dyes are not needed since the pHrodo™ Red dye is non-fluorescent outside the cell, making the staining protocol simple and fast. The elimination of wash steps and quencher dyes also improves assay reproducibility, particularly in plate reader-based assays.

Flexible applications

Use ready-made pHrodo™ Red BioParticles™ conjugates for convenient analysis of phagocytosis of Gram-positive or Gram-negative bacteria, or the amine-reactive pHrodo™ Red SE form for labeling microorganisms or proteins of your choice.

Compatible with multiple platforms
pHrodo™ Red dye conjugates can be used in plate readers, fluorescence microscopy imaging, and flow cytometry applications. The pHrodo™ Red Phagocytosis Particle Labeling Kit and the pHrodo™ Red E. coli BioParticles™ Phagocytosis Kit for flow cytometry were designed for rapid and convenient measurements of phagocytic activity in whole blood samples by flow cytometry. The kits include all the reagents required for assessing particle ingestion and red blood cell lysis. The Labeling Kit also contains the reagents required for labeling microorganisms with the pHrodo™ Red dye. Sufficient reagents are provided in both kits for approximately 100 assays.

The optimal absorption and fluorescence emission maxima of the pHrodo™ Red BioParticles™ conjugate is approximately 560 nm and 585 nm, respectively. However, the fluorophore is readily excited with the 488 nm argon-ion laser installed on most flow cytometers.