Show Order Info
SKU AR1156
Pack size 1 kit (for 500 assays)
Applications Measurement of cell proliferation in response to growth factors, cytokines and nutrients.
Measurement of cytotoxicity
To study cell activation

List of Components

DescriptionQuantityVolume ContentsCatalog Number
MTT labeling reagent15mL5mg/ml in PBSAR1156-1
Formazan solubilization solution155mL10% SDS in 0.01 M HClAR1156-2

Overview

Product Name MTT Cell Proliferation and Cytotoxicity Assay Kit
SKU/Catalog Number AR1156
Pack Size 1 kit (for 500 assays)
Storage & Expiration Upon receipt store MTT labeling reagent at -20°C. Protect from light. It is stable at -20°C for one year.
Store Formazan solubilization solution at room temperature. It is stable at room temperature for one year.
Equivalent Thermofisher (Product No. V13154), Millipore Sigma (Product No. 11 465 007 001)
For Use With (Equipment) Microplate Reader
Detection Method Colorimetric
Format 96-well plate
Label or Dye MTT
Description Boster’s MTT Cell Proliferation Assay Kit provides a simple method for determination of cell number using standard microplate absorbance readers.
Cite This Product MTT Cell Proliferation Assay Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR1156)
Application Measurement of cell proliferation in response to growth factors, cytokines and nutrients.
Measurement of cytotoxicity
To study cell activation
*Our Boster Guarantee covers the use of this product in the above tested applications.

Assay Principle

Determination of cell growth rates is widely used in the testing of drug action, cytotoxic agents and screening other biologically active compounds. Several methods can be used for such determinations, but indirect approaches using fluorescent or chromogenic indicators provide the most rapid and large scale assays. The non-radioactive, colorimetric assay system using MTT was first described by Mosmann. The assay is designed for the spectrophotometric quantification of cell growth and viability without the use of radioactive isotopes. The MTT assay involves the conversion of the water soluble MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) to an insoluble purple formazan crystals. The formazan crystals formed are then solubilized, and the concentration of resulting colored solution determined by optical density at 570 nm. The result is a sensitive assay with excellent linearity up to approximately 106 cells per well.

Important Product Information

• The culture conditions used to grow the cells can affect the results and must be taken into consideration when analyzing the data. The age of the cultures, number of passages and details of the growth medium can all be important factors.

• For Formazan solubilization solution, frozen block or precipitates may form during shipment or storage, in which case the container should be warmed to +37°C and thoroughly mixed.

• After thawing, the MTT labeling reagent may be stored protected from light at 4°C for up to 4 weeks, in which case a sterile filtration of the reagent is recommended.

• The presence of phenol red in the final assay samples can seriously affect results. We recommend that the cells be cultured in medium free of phenol red, if possible.

• MTT is oncogenic. Please wear gloves to operate.

Additional Materials Required

• 10μL, 100-200μL and multi-channel pipettes

• 96 well plate

• CO2 incubator

• DMSO

• PBS

• Plate reader capable of reading absorbance at 570nm

Assay Protocol

1. Collect cells at logarithmic phase, count the cells.
Note: Add 2000 cells per well for cell proliferation assay and 5000 cells per well for cell cytotoxicity assay. Cells number of each well depends on cell density and growth rate.

2. For adherent cells, remove the medium and replace it with 100 µL of fresh culture medium. For non-adherent cells, centrifuge the 96 well microplate, pellet the cells, carefully remove as much medium as possible and replace it with 100 µL of fresh medium.

3. Add 10 μL of the MTT labeling reagent to each well. Include a negative control of 10μL of the MTT labeling reagent added to 100 µL of medium alone.

4. Incubate the microplate at 37°C for 4 hours in a humidified chamber (5% CO2 ). At high cell densities (>100,000 cells per well) the incubation time can be shortened to 2 hours.

5. Add 100 μL of Formazan solubilization solution to each well and mix thoroughly.

6. Incubate the microplate at 37°C for 4-18 hours in a humidified chamber (5% CO2 ).

7.Check for complete solubilization of the purple formazan crystals and measure the spectropho- tometrical absorbance of the samples using a microplate reader at 570 nm.
Note: The wavelength between 560 and 600 nm can be chosen if 570nm is unavailable.

Quick Protocol Option

To shorten the time of the assay it is possible to use DMSO (not provided) as a solubilizing agent to dissolve the formazan.

1. After labeling the cells with MTT, as described above, remove all but 25 µL of medium from the wells. For non-adherent cells it may be necessary to first centrifuge the plates to sediment the cells.

2. Add 50 µL of DMSO to each well and mix thoroughly with the pipette.

3. Incubate at 37°C for 10 minutes.

4. Check for complete solubilization of the purple formazan crystals and measure the spectropho- tometrical absorbance of the samples using a microplate reader at 540 nm.

MTT Cell Proliferation and Cytotoxicity Assay Kit Images

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MTT Cell Proliferation and Cytotoxicity Assay Kit
MTT Cell Proliferation and Cytotoxicity Assay Kit
Quantitation of U937 cells using the MTT Cell Proliferation Assay Kit. Cells in the parent culture were counted in a hemacytometer and then diluted to the indicated cell numbers in 100 µL volumes, delivered to the wells of a microplate and incubated for 4 hours to allow time for adsorption before being assayed. Absorbance measurements at 570 nm were made using a microplate reader. Each data point represents the mean value of samples in triplicate.
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Publications

Rapamycin Inhibits the Growth and Collagen Production of Fibroblasts Derived from Human Urethral Scar Tissue
The effect of mast cells on the biological characteristics of prostate cancer cells
Effect of CLIC1 gene silencing on proliferation, migration, invasion and apoptosis of human gallbladder cancer cells
Long non-coding RNA PTENP1 interacts with miR-193a-3p to suppress cell migration and invasion through the PTEN pathway in hepatocellular carcinoma
Compound anisodine affects the proliferation and calcium overload of hypoxia-induced rat retinal progenitor cells and brain neural stem cells via the p-ERK1/2/HIF-1?/VEGF pathway
Differential induction of apoptosis and autophagy by pyrrolizidine alkaloid clivorine in human hepatoma Huh-7.5 cells and its toxic implication
Single-cell pH imaging and detection for pH profiling and label-free rapid identification of cancer-cells
Antioxidant axis Nrf2-keap1-ARE in inhibition of alcoholic liver fibrosis by IL-22
Nur77 attenuates endothelin-1 expression via downregulation of NF-?B and p38 MAPK in A549 cells and in an ARDS rat model
Targeting Btk with ibrutinib inhibit?gastric?carcinoma?cells?growth
Chinese herbal formula QHF inhibits liver cancer cell invasion and migration
Bortezomib inhibits cell proliferation in prostate cancer
Exogenous spermine inhibits the proliferation of human pulmonary artery smooth muscle cells caused by chemically-induced hypoxia via the suppression of the ERK1/2- and PI3K/AKT-associated pathways
Effect of ebosin on modulating interleukin-1?-induced inflammatory responses in rat fibroblast-like synoviocytes
Paxillin regulates vascular endothelial growth factor A-induced?in vitro?angiogenesis of human umbilical vein endothelial cells
5-FU and ixabepilone modify the microRNA expression profiles in MDA-MB-453 triple-negative breast cancer cells
The Jagged-2/Notch-1/Hes-1 Pathway Is Involved in Intestinal Epithelium Regeneration after Intestinal Ischemia-Reperfusion Injury
Qian YY, Li K, Liu QY, Liu ZS. Oncotarget. 2017 Nov 6;8(64):107859-107869. doi: 10.18632/oncotarget.22305. eCollection 2017 Dec 8. Long non-coding RNA PTENP1 interacts with miR-193a-3p to suppress cell migration and invasion through the PTEN pathway in hepatocellular carcinoma
Ma Z, Yue L, Xu Z, Zeng S, Ma Y, Li Z, Li W, Wang D. Cent Eur J Immunol. 2018;43(1):1-8. doi: 10.5114/ceji.2018.74867. Epub 2018 Mar 30. The effect of mast cells on the biological characteristics of prostate cancer cells
Chai L, Kang XJ, Sun ZZ, Zeng MF, Yu SR, Ding Y, Liang JQ, Li TT, Zhao J. Cancer Manag Res. 2018 May 3;10:989-1003. doi: 10.2147/CMAR.S163335. eCollection 2018. MiR-497-5p, miR-195-5p and miR-455-3p function as tumor suppressors by targeting hTERT in melanoma A375 cells
He YM, Zhang ZL, Liu QY, Xiao YS, Wei L, Xi C, Nan X. J Cell Mol Med. 2018 May;22(5):2569-2579. doi: 10.1111/jcmm.13499. Epub 2018 Mar 8. Effect of CLIC1 gene silencing on proliferation, migration, invasion and apoptosis of human gallbladder cancer cells
Yujie Jiang,1,2,* Yi Zeng,3,* Xia Huang,1,2 Yueqiu Qin,4 Weigui Luo,2 Shulin Xiang,5 Suren R. Sooranna,6 and Liao Pinhu Am J Physiol Lung Cell Mol Physiol. 2016 Dec 1; 311(6): L1023–L1035. Published online 2016 Oct 7. doi: 10.1152/ajplung.00043.2016 Nur77 attenuates endothelin-1 expression via downregulation of NF-κB and p38 MAPK in A549 cells and in an ARDS rat model

Customer Q&As

Q: Will using DMSO to solubilize the formazan crystals affect the cells since it is toxic at high concentrations?
A: DMSO will not affect the cells.