Cell Viability, Proliferation & Cytotoxicity Cell-based models continue to be critical tools for drug discovery. Assays to mea- sure viability, proliferation, and cytotoxicity are commonly used to monitor the response and health of cells in culture after treatment with various drugs, chemi- cals, or stimuli. BioVision offers an extensive range of easy-to-use, non-radioac- tive and high-throughput assays to specifically detect the number of living cells (Viability assay), the number of proliferating cells (Cell proliferation assay), and/ or the number of dead cells (Cytotoxicity assay). Many of these assays are lu- minescence-, fluorescence- or colorimetric- based, offering sensitivity, conve- nience and accuracy that correlates well with the traditional isotope-based assays. Assays to Detect Living and/or Proliferating cells Parameter Type Assay Name Incuba- Detection Sample Sensitivity Unique Features tion Time Type Suspension - High-throughput. ApoSENSOR™ Cell 10 min. Luminescence or Adherent 10-100 cells - Highly Sensitive. Viability (K254) cells - Measures ATP levels. ApoSENSOR™ Suspension ADP/ATP ratio 10 min. Luminescence or Adherent 100 cells - High-throughput. (K255) cells - Measures ADP/ATP ratio. StayBrite™ - rLucHS: highly stable & highly sensitive Highly Stable ATP Tissues & 1 nmol to 10 Luciferase. Bioluminescence 5-10 min. Luminescence Cells fmol - High-throughput. Assay (K791) - Measures ATP levels. Established ATP and/or ADP Absorbance ATP Colorimet- (OD 570 nm)/ - Does not require luminometer. Viable Cell ric/ Fluorometric 30 min. Fluorescence Tissues & 50 pmol (1 µM) - Highly stable. Biomarker Assay (K354) (Ex/Em = 535/587 nm) Cells - Measures ADP levels. Absorbance ADP Colorimet- (OD 570 nm)/ - Does not require luminometer. ric/ Fluorometric 30 min. Fluorescence Tissues & 1 µM - Highly stable. Assay (K355) (Ex/Em = 535/587 nm) Cells - Measures ADP levels. - Does not require luminometer. Absorbance - Highly stable. ADP Colorimetric 20-30 min. (OD 450 nm) Tissues & <20 µM - Measures ADP levels in samples that Assay II (K356) Cells contain reducing substances, which may interfere with oxidase-based assays EZViable™ Calcein Fluorescence Suspension Intracellular AM Cell Viability 30 min. (Ex/Em = 485/530 nm) or Adherent ~ 50 cells - High throughput. Measures only intact esterase Assay (K305) cells and live cells. - Just add & read. No washing, harvesting Quick Cell Pro- Absorbance Suspension and solubilization steps. liferation Assay 30 min.-4 hrs (OD 440 nm) or Adherent ~400 cells - Rapid and more sensitive than MTT and (WST-1) (K301) cells XTT assays. Enzymatic Ready-to-use - Ready-to-use reagent. Just add & read. Cell Proliferation Suspension No washing, harvesting and Activity Mitochondrial Colorimetric 30 min.-4 hrs Absorbance or Adherent ~400 cells solubilization steps. dehydrogenases Reagent, WST-1 (OD 440 nm) cells - Rapid and more sensitive than MTT and (K304) XTT assays. Quick Cell Prolif- - Just add & read. No washing, harvesting eration Assay plus Absorbance Suspension and solubilization steps. (proprietary WST) 30 min.-4 hrs (OD 440 nm) or Adherent ~300cells - Rapid and more sensitive than MTT, XTT (K302) cells and WST-1 assays. - Just add & read. No washing, harvesting MTS Cell Prolif- Absorbance Suspension and solubilization steps. MTS reduction eration Assay 30 min.-4 hrs (OD 490 nm) or Adherent ~600 cells - Rapid and more sensitive than MTT and (K300) cells XTT assays. General MTT Cell Prolifera- Absorbance Suspension Metabolic MTT reduction tion Assay (K299) 3 hrs (OD 590 nm) or Adherent ~ 900 cells Convential assay to measure cell proliferation Activity cells Fluorescence - Just add & read. No washing, harvesting Vision Blue™ Vision Blue™ (Ex/Em = 530-570 nm/ Suspension and solubilization steps. reduction Quick Cell Viabil- 1-5 hrs 590-620 nm) or Adherent ~100 cells - Rapid and more sensitive than MTT, XTT, ity Assay (K303) cells WST-1 and proprietary WST assays. 155 S. Milpitas Blvd, Milpitas, CA 95035 T: 408-493-1800 F: 408-493-1801 Toll Free: 800-891-9699 (US Only) Parameter Type Assay Name Incubation Detection Sample Sensitivity Unique Features Time Type BrdU Cell Pro- Suspension ~50-100 Brdu liferation Assay 4-8 hrs Absorbance or Adherent proliferating - Only measures the proliferating, viable incorporation (K306) (OD 450 nm) cells cells cells. DNA - Just add-and-read. No washing, Synthesis Cell Proliferation Fluorescence (Ex/Em = Suspension harvesting, and solubilization steps. Nuclear Dye 15 min. or Adherent 25 cells - Non-radioactive. Assay (K307) 480/538 nm) cells - High-throughput method for cell proliferation and apoptosis. Assays to Detect Dead Cells Parameter Type Assay Name Incubation Detection Sample Sensitivity Unique Features Time Type LDH-Cytotoxicity 30 min.-1 hr Absorbance Suspension ~200-250 cells - Stable and abundant enzyme. Assay (K311) (OD 495 nm) or Adherent - Based on reaction of pyruvate with cells tetrazolium salt INT to form formazan. LDH-Cytotoxicity < 1 hr Absorbance Suspension ~500-600 - Minimize background from sreum & culture Assay II (K313) (OD 450 nm) or Adherent cells media: culture cells in regular 10% serum. Lactate De- cells - More stable: stop reaction at any time point hydrogenase and read multiple times. Enzymatic released from - Based on reaction of NADH with WST to compromised generate color. Activity (leaky) cells PicoProbe™ < 20 min. Fluorescence Suspension ~100 cells - High sensitivity LDH-Cytotoxicity (Ex/Em = 535/587 nm) or Adherent - High-throughput Assay (K314) cells - Stable Enzyme Adenylate Bioluminescence 30 min. Luminescence Suspension ~50-100 cells - High throughput. kinase released Cytotoxicity or Adherent - Simple one step procedue involving two from compro- Assay (K312) cells chemical reactions mised (leaky) cells Cell-permeable Live-Dead Cell 20 min. Fluorescence Suspension Single Cell - Visualize and/or sort the live and dead cells. green Staining Kit (K501) Microscopy/Flow or Adherent - Analyze the apoptotic cells. fluorescent Cytometry cells dye: To Live/Dead label live cells Cell Fluorescent Cell nonperme- Cell-Mediated 30 min. Fluorescence Suspension Single Cell - Measure cell-mediated cytotoxicity. able Cytotoxicity Microscopy/Flow or Adherent - Visualize and/or sort the live and dead cells. Probes red Fluorometric Cytometry cells - Analyze apoptotic/necrotic cells. fluorescent dye: Assay Kit (7-AAD/ - Broader applications. to label late CFSE) (K315) - Multi-parametric analysis. apoptotic and necrotic cells www.BioVision.com [email protected] 2.