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CBA410 EZMTTTM Cell Proliferation Assay, MTT based, 200X

A novel tetrazolium salt based proliferation assay for continous monitoring of growth in mammalian and bacterial cell culture.

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CBA410
0.5 mL  
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      Overview

      Replacement Information
      Description
      Catalogue NumberCBA410
      DescriptionEZMTTTM Cell Proliferation Assay, MTT based, 200X
      OverviewNAD+/NADH and NADP+/NADPH are important biological cofactors for oxidoreductase enzymes that are fundamental for various biological processes, such as energy metabolism, mitochondrial function, oxidative stress, immunological functions and cell death. Hence the oxidoreductase enzymes are used as indicators of metabolic activity of cells.

      EZMTT is a novel monosulfonated tetrazolium salt, 2-(3-(2-methoxy-4-nitrophenyl)-2-(4-nitrophenyl)-2H-tetrazol-3-ium-5-yl) benzenesulfonate sodium salt (EZMTT). The EZMTT reagents provided high quality and reproducible detection for oxidoreductase enzymes, indicators of cellular metabolic activity. Upon reacting with metabolic enzymes, EZMTT is converted to a yellowish formazan that does not require a solubilization step and can be measured by absorbance at 450 nm. EZMTT assay can continuously track cell proliferation and measure cytotoxicity (loss of viable cells) or cytostatic activity (shift from proliferation to quiescence) of potential medicinal or toxic agents.

      Traditional tetrazolium salts, such as MTT, react with NAD(P)H-dependent cellular oxidoreductase enzymes to produce an insoluble formazan, which requires solubilization, has lower sensitivity, and is an end-point assay.

      Features and Benefits of EZMTT:
      • Can be premixed with any media and then directly added to the cells
      • Is very stable and has little cytotoxicity. Incubation time depends on the cell number tested, if the color difference in cultures with/without cells is not obvious, a longer incubation from hours to weeks is possible.
      • Reduction of EZMTT produces a water-soluble yellow colored formazan dye, which does not require solubilization step
      • More sensitive than the traditional MTT assays and can track the cell growth easily (mammalian as well as bacterial cells) by absorbance at 450 nm or color changes. With longer incubation time, as little as 1 bacteria cell can be detected.
      • Much less reactive with other antioxidants such as beta-Mercaptoethanol (BME)

      Assay Protocol
      1. Detach Hela cells and seed different cell number (0, 2500, 5000, 10000, 20000, 30000, 40000, 50000) in each well of 96-well plate with 10% FBS DMEM (100 μl) in triplicates.
      2. Incubate at 37⁰C, 5% CO₂ for 3-4 hours or overnight.
      3. Thaw and Mix 200 X EZMTT cell proliferation assay solution with culture media (20-fold dilution).
      4. Add 10 μl of the 10X EZMTT solution to each well of the plate (final 200-fold dilution).
      5. Incubate the plate for 1-4 hours in the incubator, measure the absorbance at 450 nm using a microplate reader.

      References
      1. Zhang W, et al. Mono-sulfonated tetrazolium salt-based NAD(P)H detection reagents
      suitable for dehydrogenase and real-time cell viability assays. 2016. Anal Biochem. 509, 15 Sept 2016, 33-40.
      2. Hu Q, et al. Detection of "Hidden" Antimicrobial Drug Resistance. 2019. ACS Infectious Diseases. 5, 1252-1263
      3. Ruan J, et al Kidney-Type Glutaminase Inhibitor Hexylselen Selectively Kills Cancer Cells via a Three-Pronged Mechanism, 2019. ACS Pharmacology & Translational Science, 2019, 2: 18-30.
      4. Rui J, et al. The EZMTT cell proliferation assay provides precise measurement for drug combinations and better correlation between in vitro and in vivo efficacy. 2020. Bioorganic & Medicinal Chemistry Letters 30,127134-127239
      Alternate Names
      • MTT assay, Proliferation assay
      • Viability assay
      References
      Product Information
      Detection methodColorimetric
      PresentationSolution
      Applications
      ApplicationA novel tetrazolium salt based proliferation assay for continous monitoring of growth in mammalian and bacterial cell culture.
      Key Applications
      • in vitro Toxicology Assays
      • Cell Based Assays
      Application NotesCell Proliferation, Cell viability and Cytotoxicity assays
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssurancePurity: ≥ 98% confirmed by TLC or NMR. Performance tested by NADH titration of product, dose response to E. coli culture, and dose response to A549 cells.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStore EZMTT cell proliferation assay at -20°C and protect from light.
      Packaging Information
      Material Size0.5 mL
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      CBA410 04061841829812