Cell Health Assays Selection Guide

Quickly Find the Ideal Assay for Cell Health Analysis

Molecular Probes offers a diverse selection of fluorescence based assays for the analysis of apoptosis, cell cycle, cell viability and other aspects of cell health. Many of these technologies can be used across multiple platforms. In an effort to simplify the decision process, we've developed this assay selection guide. Simply click on a parameter of cell health under the chosen platform and you will be shown an ideal product for that type of assay.

Download the Cell Health Selection Guide [PDF, 1.75MB]

Imaging Assays

Apoptosis
Autophagy
Cell Cycle
Cell Proliferation
Cell Tracking
Cell Viability
Cytoskeletal Changes
Endocytosis & Phagocytosis
GFP/RFP Detection
Oxidative Stress
Protein Detection
RNA Detection

HCS Assays

Cell Proliferation
DNA Damage Analysis
Lipid Detection
Cell Viability
Mitotoxicity & Cytotoxicity
New Protein Synthesis
Proliferation, Cell Division (Mitosis)
RNA Synthesis

Flow Assays

Apoptosis
Cell Cycle
Cell Proliferation
Cell Viability

Microplate Assays

Autophagy
Cell Proliferation
Cell Viability

Click-iT® TUNEL Alexa Fluor® Imaging Assay

Cat. No. C10245, C10246, C10247

BASIC PRINCIPLE: Detection of fragmented DNA during Apoptosis, by Click iT® TUNEL assay

• Robust workflow can be completed in 3 hours
• Available with Alexa Fluor® 488, 594 and 647
• Much more sensitive than other TUNEL methods
• Multiplexible with most other fluorescent reagents
• Ideal for Imaging in vivo and in vitro, compatible with HCS and microplate assays

CellEvent® Caspase-3/7 Green Detection Reagent

Cat. No. C10423

BASIC PRINCIPLE: Caspase 3/7 activity during apoptosis

• Simple, 30 minute no-wash protocol
• Absorption/emission maximum: ~502/530 nm
• Suitable for live cell fluorescence-imaging, enabling continuous monitoring of dynamic events
• Fixable and permeable
• DEVD peptide sequence conjugated to a nucleic acid binding dye
• Ideal for Imaging, HCA/HCS, HTS, and flow cytometry

Autophagy Detection Reagents

Cat. No. P36235, P36236, L10382

BASIC PRINCIPLE: Reagents based on localization of LC3B protein to autophagosomes

• Specific tools to report LC3B localization: an antibody against LC3B, and Premo™ sensors
• High efficiency transduction: The Premo™ Autophagy Sensor LC3B leverages BacMam 2.0 delivery technology
• GFP or RFP constructs: The Premo™Autophagy Sensors are fused to the
N-term of LC3B, with the C-term free for cleavage and recruitment of autophagosomes
• Image the localization of cellular LC3B using a highly specific LC3B antibody

Premo™ FUCCI Cell Cycle Sensor, BacMam 2.0

Cat. No. P36237

BASIC PRINCIPLE: Cell cycle sensor that changes from green to red to green with cell cycle phases.

• Live cell indicator of cell cycle status. Red or green fluorescence indicates stage of cell cycle
• Flexible: works with live cells, but compatible with fixation
• Convenient: Uses BacMam 2.0 delivery system: Add to cells, incubate overnight, visualize the next day
• Enables cell cycle analysis in a majority of adherent cell types (cell lines, primary cells and stem cells) with high efficiency and exceptional workflow - add, incubate and read

Click-iT® EdU Kits

Cat. No. C10337, C10339, C10340, C10338

BASIC PRINCIPLE: Integration and detection of EdU in nascent DNA, during proliferation

• Easy protocol
• Typically takes 2 hours
• Sample left intact
• Highly sensitive and accurate assay that leaves cell morphology intact

Multiple Formats
• Imaging, Flow, HCS, Microplate (HCS compatible)

CellTracker™ Dyes in various colors

Cat. No. C7025, C34552, C34551, C7000, C2110,C10094,C12881,C2925,C2102,C2927, C2111

BASIC PRINCIPLE: Small dye based fluorescent probes to label live cells, which can be traced for several generations

• Non-Toxic: No known effect on proliferative ability or biology of cells
• Easy to label cells: trackers pass freely through cell membranes
• Long-term signal stability: Well retained in cells for several days post staining
• Fixable: compatible with fixed cell analysis and long-term storage
• Multiple colors available : blue, violet, green, orange and red

Image-iT® DEAD Green™ Viability Stain

Cat. No. I10291

BASIC PRINCIPLE: A highly sensitive non-fluorescent dead cell stain that fluoresces brightly green on binding to DNA in cells with damaged cell membranes

• Clear results: Bright green fluorescence by membrane impermeant dye indicates cytotoxicity and membrane damage
• Multiplexible: compatible with blue and red dyes
• Flexible: Survives fixation and permeabilization for combination with additional parameters of cytotoxicity or other biomarkers. Compatible for traditional fluorescent microscopy and HCS
• Sensitive: Excellent discrimination of live and dead cells

Alexa Fluor® Phalloidin conjugates

Cat. No. A12379,R415,A12381, A12380,A22287, A22283, A34055

BASIC PRINCIPLE: Phallodin is a peptide with high affinity for the actin filament. Fluorescent phalloidin conjugates provide highly sensitive probes to image cytoskeleton and cell morphology.

• Selective and specific: Selectively stains F-actin (key cytoskeletal component) more specifically than antibody methods
• Alexa Fluor® dyes give exceptional brightness, stability, and broad color options
• For fixed & permeabilized cells
• Validated in thousands of papers: 3,000 on cancer alone!

pHrodo™ pH sensors

Cat. No. P36600,P35361,P10361, A10010

BASIC PRINCIPLE: Bright red fluorescence in lower pH environmentsuch as phagocytic or endocytic vesicles

• Fluorescent detection of phagocytosis and endocytosis—discriminate endocytosis from adherent and extracellular particles
• No wash or quencher dyes required: Reduce signal variability and improve timing in sensitive experiments
• Multiplexible: bright red fluorescence is conveniently multiplexible with green dyes such as GFP, fluo-4, or calcein
• Available as convenient conjugates: Dextran and bioparticle conjugates available, also available as labeling reagent (SE form)

Anti-GFP and RFP antibodies

Cat. No. A6455,A10262,A11122, A11120, A11121, G10362, R10367, A21311, A31851, A21312, A31852

BASIC PRINCIPLE: Antibodies to detect GFP and RFP

• Sensitive and specific detection: Alexa Fluor® dye– conjugated and purified antibodies offer optimal specificity and low background
• Ready for any platform: choose from a variety of hosts, isotypes, and applications
• Rabbit monoclonal anti-GFP antibody, produced with exclusive ABfinity™ technology

CellROX® Deep Red Reagent

Cat. No. C10423

BASIC PRINCIPLE: Upon oxidation by ROS exhibits deep red fluorescence

• General reactive oxygen species (ROS) or oxidative stress detector
• Compatible with fluorescent imaging, florescent plate readers, flow cytometry, and HCA/HCS
• Highly resistant to photobleaching
• Multiplexible: GFP and Alexa Fluor® 488 compatible (EX/EM: ~644/665nm)
• Versatile: Suitable for live/fixed-cell imaging

Click-iT® AHA and HPG Nascent Protein Synthesis

Cat. No. C10102, C10186, A32761, A32762, A32763

BASIC PRINCIPLE: Incorporation and detection of Click-iT® AHA or HPG in nascent protein

• Novel: unique approach to study protein synthesis, stability and kineticsincorporation of amino acids Click-iT® AHA (L-azidohomoalanine) or Click-iT® (L-homopropargylglycine) into nascent polypeptide chains
• Less hazardous: eliminate need for radioactivity
• Flexible: multiplex with antibody and dye-based methods
• Sensitive: efficient click method and bright Alexa Fluor® dyes help ensure high sensitivity

Click-iT®-Based Detection of Nascent RNA Synthesis

Cat. No. C10329, C10330

BASIC PRINCIPLE: Incorporation and detection of Click-iT® EU in Nascent RNA

• Physiologically relevant: Nucleoside analog EU (5- ethynyl uridine
fed to cells and incorporated into nascent RNA
• Novel: unique approach to study mRNA turnover, stability and kinetics
• Safe alternative: eliminate radioactivity
• Flexible: multiplex with antibody and dye-based methods
• Sensitive: efficient, proven Click-iT® method combined with bright Alexa Fluor® dyes help ensure high sensitivity
• Economical: eliminate compounds with off-target effects early in discovery process

Click-iT® EdU HCS Cell Proliferation Assays

Cat. No. C10357, C10337, C10340, C10339, C10355, C10350, C10356, C10351, C10354, C10352, C10353

BASIC PRINCIPLE: Assays use a modified nucleoside, EdU, that is incorporated during DNA synthesis and detected with a quick and highly specific click chemistry reaction

• Accurate: superior to BrdU assays with minimal variation (low CVs) and much simpler protocol
• Speed: results in as little as 90 minutes
• Sensitivity: no DNA degradation and super bright Alexa Fluor® labels afford outstanding sensitivity
• Flexibility: easily multiplex with dye and antibody based imaging reagents

HCS DNA Damage Kit

Cat. No. H10292

BASIC PRINCIPLE: Simple, fast and robust assay for doublestranded DNA breaks. Phospho-H2AX is a sensitive marker for DNA damage and repair

• Economical: two assays in one
• Accuracy: simultaneous detection of genotoxicity (DNA damage /phosphorylated H2AX - red) & plasma membrane integrity (cytotoxicity - green) helps improve accuracy
• Confidence: two parameter cytotoxicity assay minimizes risk of false negatives
• Established: common readout in cytotoxicity and cancer research

HCS LipidTOX™ Phospholipidosis and Steatosis

Cat. No. H34157, H34158, H34350, H34351, H34475, H34476, H34477

BASIC PRINCIPLE: The kit offers a complete set of reagents for performing image-based high-content assays to characterize phospolipidosis and steatosis, toxic side effects on lipid metabolism that can be triggered by drugs and other compounds

• Relevant: Labeled phospholipids incorporated into live cells
• Economical: Cost and time saving—detect and remove compounds with side effects early in discovery process
• Convenient: Ready-to-use aqueous formulations for easy assay preps. No sonication or organic solvent required, fast—fixed endpoint workflow
• Flexible: Green and red emission choices with tight peak spectra for multiplexing

HCS LIVE/DEAD® Green Kit

Cat. No. H10290

BASIC PRINCIPLE: Measures cytotoxicity by membrane permeability and nuclear morphology. Cell injuries from toxic compounds include membrane damage, allowing entry of the Image-iT® DEAD Green™ viability stain. Total (live and dying) cell numbers and nuclear morphology is measured by either blue (Hoechst 33342) or far red (HCS NuclearMask™ Deep Red) stains.

• Highly sensitive cytotoxicity assay that also discriminates live and dead cells
• Fast and efficient: Dye-based, two color assay with convenient workflow
• Multiplexible: Two color choices for live cell staining for multiplex flexibility
• Flexible: Compatible with fixation/ permeabilization methods for combination with antibody-based assays

HCS Mitochondrial Health Kit

Cat. No. H10295

BASIC PRINCIPLE: Multiplex assay for simultaneous detection and quantitation of pre-lethal (mitotoxicity) and lethal (cytotoxicity) cell damage

• Economical: three assays in one
• Convenient: dye-based assays are faster than antibody-based methods
• Compatible with live cell analyses
• Higher data quality: simultaneous detection of three parameters improves data quality and reduces risk of false negatives
• Physiologically relevant: combination of mitotoxicity, cytotoxicity and morphological readout enhances correlation to in vivo models)

Click-iT® AHA Alexa Fluor® 488 Protein Synthesis HCS Assay

Cat. No. C10289

BASIC PRINCIPLE: Fast, sensitive, non-toxic, and non-radioactive method for imagebased detection of new protein synthesis. Enabling method for analysis of protein expression or turnover in disease models, drug discovery or from environmental changes

• Unique assay—Click-iT® kit for new protein detection and protein synthesis, stability and kinetics

• Simple—based on incorporation of “clickable” amino acid
• Less hazardous
• Flexible—multiplex with antibody and dye-based methods
• Sensitive—efficient click method and bright Alexa Fluor® dyes help ensure high sensitivity and accuracy
• Saves time and money—get relevant answers faster and in drug discovery eliminate compounds with off-target effects early in discovery process

HCS Mitotic Index Kit

Cat. No. H10293

BASIC PRINCIPLE: Multiplex assay to quantify dividing (mitotic) cells and total DNA content. Many anti-cancer drugs target the cell cycle and stop cell division.

• Accurate—Phospho histone 3 is a known mitotic marker, commonly used in cancer research, toxicology and cell cycle signaling
• Specific—Anti histone H3 Alexa Fluor® 488 Ab indicates mitotic arrest
• Flexible—Use DAPI or NuclearMask™ Deep Red for total DNA content
• Excellent assay robustness with good CV values and Z’ scores

Click-iT® RNA Alexa Fluor® HCS Kit

Cat. No. C10327, C10328

BASIC PRINCIPLE: Click-iT®-based detection of RNA synthesis in kit optimized for high-content analysis. Enables the study of mRNA stability and turnover kinetics in normal and disease-related models (e.g. autophagy, cancer). Same labeling principle as Click-iT® EdU.

• Unique assay—Click-iT® kit for RNA detection
• Non-radioactive—fluorescent alternative
• Flexible—multiplex with antibody and dye-based methods
• Sensitive—efficient click method and bright Alexa Fluor® dyes help ensure high sensitivity
• Saves time and money—eliminate compounds with off-target effects early in discovery process

Violet Membrane Asymmetry Probe

Cat. No. A35137

BASIC PRINCIPLE: Two-color ratiometric response to variations in surface charge

• Novel—Accurate apoptotic analysis on trypsinized cells (adherent cells)
• Fast—Simple 5-minute staining protocol
• Compatible—Easily combined with other blue-excited apoptotic stains

Vybrant® DyeCycle™ Stains

Cat. No. V35003, V35004, V35005, V10273, V10309

BASIC PRINCIPLE: Cell membrane permeant nucleic acid stains that emit a fluorescence signal proportional to DNA mass

• Precise—Enables accurate cell cycle analysis in living cells

• Safe—Low cytotoxicity for cell sorting and additional live cell experiments
• Flexible—Multiple colors available, for all common laser lines

CellTrace™ Violet and CFSE

Cat. No. C34554, C34557

BASIC PRINCIPLE: Dye dilution through cell division

• Superior performance—Bright, single peak staining enables visualization of multiple generations
• Long-term signal stability—Well retained in cells for several days post stain
• Non-cytotoxic—No known effect on proliferative ability or biology of cells
• Versatile—Multiple colors available to easily combine with antibodies or markers of cell function, such as GFP
• Novel—multiple vendors for CSFE, but significant advantages for CellTrace Violet make it a better choice

Click-iT® EdU Cell Proliferation Assays

Cat. No. C10418, C10419, C10424, C10425, C10420

BASIC PRINCIPLE: Assays use a modified nucleoside, EdU, that is incorporated during DNA synthesis and detected with a quick click chemistry reaction

• Accurate—superior to BrdU assays with minimal variation (low CVs) and much simpler protocol
• Fast—results in as little as 90 minutes
• Simplicity—streamlined protocol with fewer components than the original kits

LIVE⁄DEAD® Fixable Dead Cell Stains

Cat. No. L23105, L34955, L34957, L34959, L23101, L23102, L10120, L10119

BASIC PRINCIPLE: Cell membrane integrity and fluorescent intensity

• Fixable—Staining retained after fixation for simple live/dead analysis with intracellular phenotyping
• Simple—fits into almost any staining and phenotyping protocols
• Flexible—seven colors from which to choose for UV, 405, 488, or 633 lasers

Sample Pack recommended for panel optimization

Order LIVE/DEAD Sample Pack now

SYTOX® Dead Cell Stains

Cat. No. S10274, S10349, S34861, S34857, S34859, S34860, S34862

BASIC PRINCIPLE: Membrane impermeable nucleic acid stains

• Accurate—high-affinity nucleic acid stains for easy dead-cell discrimination
• Flexible—multiple colors with minimal spectral overlap for expanded multicolor capabilities
• Easy to use—no wash steps. Simply add, incubate, and analyze

Order SYTOX® Dead Cell Stains Sample Pack now

Premo™ Autophagy

Cat. No. A14070, A14294

BASIC PRINCIPLE: Quantitates macroautophagy in a TR-FRET assay format. The assay is portable across cell types or use with existing GFP-tagged LC3B Cell Line.

• Quantitate autophagy results
• Simple add and read format minimizes the numerous sample preparation steps required by cell staining or western blot
• Measure autophagy in physiologically relevant cell lines
• The plate-based assay enables simple handling of many samples and conditions

CyQUANT® Direct Cell Proliferation Assay

Cat. No. C35011, C35012

BASIC PRINCIPLE: Quantitates number of cells in a population based on viable cell content by measuring intensity of dyes that fluoresce upon DNA

• Measures proliferation independent of metabolic state of cells on standard fluorescent plate readers
• Convenient workflow for high-throughput screening –no washes, cell lysis, temperature equilibrations, or radioactivity required
• Rapid protocol, high sensitivity and dynamic range
• Multiplex assays using a spectrally distinct fluorescent or a luminescent readout

PrestoBlue® Cell Viability Assay

Cat. No. A13261, A13262

BASIC PRINCIPLE: Resazurin-based assay, reduced in living cells, turning from blue to red becoming highly fluorescent

• Save time with a shorter incubation step compared to other resazurinbased assays
• High-quality results with large assay window and Z’ values
• Convenient add-and-read homogenous assay format
• Monitor living cells and perform downstream functional assays with no cell lysis