MTT Cell Proliferation and Viability Assay Kit

MTT Cell Proliferation and Viability Assay Kit - Cat Number: 6034 From Chondrex.

Research Field: Cell Biology

Clonality: N/A

Cross-Reactivity:

Host Origin: N/A

Applications: N/A

Isotype: N/A

Detection Range: 1000 cells/well - 100000 cells/well

Sample Type: Cultured Cells

Concentration: N/A

Immunogen:

DESCRIPTION: Assay kit to evaluate cell viability and cell density

FORMAT: 96-well ELISA Plate with non-removeable strips
ASSAY TYPE: Colorimetric Assay
ASSAY TIME: 6+ hours
STANDARD RANGE: 1,000 to 100,000 cells/well
NUMBER OF SAMPLES: Up to 250 (duplicate) samples/kit
SAMPLE TYPES: Cultured Cells
RECOMMENDED SAMPLE DILUTIONS: N/A
CHROMOGEN: N/A (read at 570 nm)
STORAGE: -20°C
VALIDATION DATA: N/A
NOTES: This kit has an optional overnight incubation step

INTRODUCTION

Cell proliferation and viability assays are important, routine techniques in cell biology. These assays are used to evaluate biologically active compounds and drugs and cytotoxic agents, as the assay results correlate with indicators of cellular health, such as intactness of cell membrane and metabolism. Although there are several assay methods to evaluate cell viability using metabolic activities (ATP or substrates) or DNA synthesis (Thymidine or BrdU incorporation), here, Chondrex, Inc. introduces a tetrazolium reduction assay to evaluate cellular metabolic activity. Several tetrazolium compounds are available as substrates such as MTT, MTS, XTT, and WST-1.

Among these compounds, only MTT (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) is a positively charged compound which can penetrate viable eukaryotic cells and reflect cytosolic metabolic activities. Soluble MTT is taken into cells and is reduced by NADH or other reducing molecules by electron transfer. The reduced MTT forms insoluble formazan which accumulates both inside of and on the surface of cells. Generation of formazan reflects viable cell density because dead cells lack the metabolic activity to turn MTT into formazan (1). The resulting formazan can be solubilized and quantified to reflect cell proliferation and viability. Chondrex, Inc. provides an MTT cell proliferation assay kit (Cat # 6034) which employs a simple method to assay cell viability and cell density using an absorbance microplate reader. This kit can be used for up to 250 samples in duplicate.

KIT COMPONENTS

NOTES BEFORE USING ASSAY

NOTE: It is recommended that the standard and samples be run in duplicate.

ASSAY OUTLINE

ASSAY PROCEDURE

1. Prepare the cell suspension in culture media at 100 µl/well in 96-well plates.
NOTE: 1 x 103
to 105 cells per well may be an appropriate amount, however, optimization is required.
2. Incubate cells with test reagents for desired period of exposure.
3. Add 10 µl MTT Solution per well.
4. Incubate for 2 - 4 hours at 37°C.
5. Add 100 µl of Solubilizing Solution to each well.
6. Incubate the plate in a dark place at room temperature for 4 hours or overnight.
7. Read absorbance at 570 nm.

CALCULATING RESULTS

1. Average the duplicate OD values for all wells
2. Subtract the averaged OD values of the media-only wells from the averaged OD values of all of the test wells.
3. Compare the OD values between the control group and test group.
NOTE: OD values depend on cell type, cell density, and biological activity