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Cell Meter™ Colorimetric MTT Cell Proliferation Kit

Cell numbers were determined with Cell Meter™ Colorimetric MTT Cell proliferation Kit. HeLa cells at 0 to 40,000 cells/well/100 ?L were added in a clear bottom 96-well plate for overnight. The absorbance was measured at 560 nm using a SpectraMax reader (Molecular Devices). 500 cells/well was detected compare to ~5,000 cells/well with Sigma’s MTT assay kit.
Cell numbers were determined with Cell Meter™ Colorimetric MTT Cell proliferation Kit. HeLa cells at 0 to 40,000 cells/well/100 ?L were added in a clear bottom 96-well plate for overnight. The absorbance was measured at 560 nm using a SpectraMax reader (Molecular Devices). 500 cells/well was detected compare to ~5,000 cells/well with Sigma’s MTT assay kit.
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Catalog Number22768
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Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
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H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Cell Meter™ assay kits are a set of tools for monitoring cell viability. A variety of parameters can be used to monitor cell viability. Cell Meter™ Colorimetric MTT Cell Proliferation Kit uses MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) to quantify the number of live cells. The water-soluble MTT produces water-insoluble purple formazan in metabolically active cells. The amount of formazan produced is directly proportional to the number of living cells. The absorbance increase is measured around 560 nm. Unlike other commercial MTT assays that require the tedious solubilization step to dissolve the water-insoluble formazan product, Cell Meter™ Colorimetric MTT Cell Proliferation Kit is a simple mix and read format with the minimal hands-one time. Our proprietary formulation eliminates the time-consuming solubilization step. It is the most robust MTT test for determining the number of viable cells in cell proliferation and cytotoxicity assays. The detection sensitivity is 10 times higher than the traditional MTT assays.

Platform


Absorbance microplate reader

Absorbance562 nm
Recommended plateClear bottom

Components


Component A: MTT Reagent A40 mL
Component B: MTT Reagent B100 mL

Example protocol


AT A GLANCE

Protocol summary
  1. Prepare cells in a 96-well plate (100 µL/well)
  2. Add MTT working solution (140 µL/well)
  3. Incubate at 37 °C for 2-4 hours
  4. Read absorbance at 560 nm 

Important
Thaw all the kit components at room temperature before use.

PREPARATION OF WORKING SOLUTION

MTT working solution
Prepare the amount of MTT working solution needed by mixing 4 mL of MTT Reagent A (Component A) with 10 mL of MTT Reagent B (Component B) (2:5, v/v ratio of MTT Reagent A: B). Mix well.
Note     14 mL MTT working solution is enough for 100 tests in a 96-well plate. Prepare enough MTT working solution right before the experiment, use promptly.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Plate 1000 to 40,000 cells/well in a tissue culture microplate with clear bottom.
  2. Add test compounds into the cells and incubate for a desired period of time (such as 24, 48 or 96 hours) in a 37 °C,  5% CO2 incubator. For blank wells (medium without the cells), add the same amount of test compounds. The suggested total volume is 100 µL for a 96-well plate, and 25 µL for a 384-well plate.
    Note     Each cell line should be evaluated on an individual basis to determine the optimal cell density for proliferation or cytotoxicity induction. For proliferation assays, use fewer cells; for cytotoxicity assays, use more cells to start with.
  3. Add 140 µL/well (96-well plate) or 35 µL/well (384-well plate) of MTT working solution to each well.
  4. Incubate the plate at 37 °C for 2-4 hours, protect from light.
    Note     The incubation time could be from 2 hours to overnight depending on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.
  5. Monitor the absorbance increase with an absorbance plate reader at OD = 562 nm. 

References


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Journal: Environmental toxicology and pharmacology (2020): 103409
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Journal: Journal of ethnopharmacology (2019): 112-118
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Ultrasound-Stimulated Microbubbles Enhance Radiosensitization of Nasopharyngeal Carcinoma.
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