NUMER JEDNOSTKI MAGAZYNOWEJ: AFG-BV-0468

AffiASSAY® MTS Cell Proliferation Colorimetric Assay Kit

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AffiASSAY® MTS Cell Proliferation Colorimetric Assay Kit

One-step colorimetric quantification of viable cells for proliferation, metabolic activity and cytotoxicity studies.

Catalog numbers AFG-BV-0468–AFG-BV-0472
490 nm Detection wavelength Soluble formazan is quantified by absorbance.
30 min–4 h Reaction time Incubation time after addition of the MTS reagent.
One step Assay format No separate formazan-solubilization step is required.

Introduction and assay principle

MTS offers distinct advantages as a detection reagent for cell proliferation assays. It provides a direct measurement of mitochondrial metabolic activity and therefore supports the assessment of cell viability and growth.

Compared with MTT, the MTS method provides a quicker, more sensitive and easier-to-use protocol, with improved linearity. The colored formazan product generated from MTS remains soluble, eliminating the need for a separate formazan-solubilization step.

The AffiASSAY® MTS Cell Proliferation Colorimetric Assay Kit is a one-step optimized assay for quantification of viable cells in proliferation and cytotoxicity studies. Reduction of MTS by viable cells to colored formazan is believed to be mediated by mitochondrial dehydrogenases. The resulting formazan is quantified by absorbance at 490 nm.

Applications

Cell proliferation

Study cell proliferation and responses to growth factors, cytokines, mitogens and nutrients.

Cytotoxicity

Evaluate cytotoxicity caused by anticancer drugs and other potentially toxic compounds.

Metabolic inhibition

Study inhibitors that reduce cell growth or cellular metabolic activity.

Available kit sizes

Catalog number Kit size MTS reagent volume Detection method
AFG-BV-0468 250 wells 5 mL Colorimetric
AFG-BV-0469 500 wells 10 mL Colorimetric
AFG-BV-0470 2,500 wells 50 mL Colorimetric
AFG-BV-0472 5,000 wells 100 mL Colorimetric
AFG-BV-0471 10,000 wells 200 mL Colorimetric

Storage and handling

−20°C
Store the unopened kit at −20°C. For long-term storage, maintain the kit at −20°C.
Up to 6 weeks
When used regularly, the MTS reagent may be stored at 4°C for up to six weeks.
Light sensitive
Bring the reagent to room temperature before use and work under subdued light whenever possible.

Assay protocol

  1. Seed and culture the cells
    Culture cells at an appropriate density for the cell type being used. A range of 5–100 × 103 cells per well in 200 µL of culture medium, with or without the compounds to be tested, is an appropriate starting range.
  2. Incubate
    Incubate the cells for 24–48 hours.
  3. Add MTS reagent
    Add 20 µL of MTS reagent to each well containing 200 µL of culture medium. Incubate for 30 minutes to 4 hours under the same culture conditions used during the previous incubation.
  4. Measure absorbance
    Shake the plate briefly and measure absorbance at 490 nm.

Important notes

Alternative culture volumes

When cells are cultured in a different volume of culture medium, maintain the volume of MTS reagent at 10% of the culture-medium volume.

Delayed absorbance measurement

Absorbance measurement may be postponed for up to 18 hours. Add 10 µL of 10% SDS to each well to kill the cells and stop the reaction. Protect the SDS-treated plate from light at room temperature until measurement.

Representative linear response

Linear response range: 10–100 × 10³ cells per well 0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 0 20 40 60 80 100 Cell number per well (× 10³) OD 490 nm

Jurkat cell response

Jurkat cells were diluted to the indicated numbers per well in serum-free medium. MTS reagent was added and the cells were incubated at 37°C for one hour.

The plate was then measured using a plate reader at 490 nm. The response was highly linear between 10 and 100 × 103 cells per well.

y = 0.0275x + 0.3974 R2 = 0.9998
For research use only Not for use in diagnostic procedures or on humans.