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CAS:298-93-1
Appearance:Light yellow to yellow solid
Storage:Powder : 2-8℃, 2 years; In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:≥98%
Manual Download
MTT is a colorimetric agent widely used to measure cell proliferation. MTT can be reduced by some dehydrogenases in the mitochondria to produce the crystalline dark purple product formazan, which can be completely dissolved by DMSO, and then the absorbance near the wavelength of 490nm can be measured by enzymoleter. The more cells proliferate and the faster, the higher the absorbance; The greater the cytotoxicity, the lower the absorbance. Note: MTT solution should be kept away from light, long time light will cause failure. Do not use when the color turns grayish green.
Instructions for use (for reference only)
1. Collect logarithmic cells and culture cells according to their own experimental needs.
2. Carefully absorb the supernatant, add 90μL fresh medium, then add 10μL MTT solution, and continue to culture for 4 h.
3. Then absorb the supernatant, add 110μL DMSO to each hole, and oscillate at a low speed for 10 min on a shaking table to fully dissolve the crystal.
4. The light absorption value of each hole was measured at 490 nm by ELISA. Note: The commonly used concentration of MTT solution is 0.5% (for reference only), and the experimenters can also adjust it according to their own experimental needs.
Examples of using this product（for reference only）
In Vitro:
Cell （HEK-293 cells，37 ℃ ，4 h，490nm）:
Cell viability was determined by a quantitative colorimetric assay with MTT. To screen the pre-protection of SDAP1 and SDAP2, cells were pretreated with SDAP1 and SDAP2 (0.25, 0.5, 0.75, 1, 1.25, 1.5 mg/mL) before administration of GM (3 mg/mL) for 24 h. HEK-293 cells were dispensed in 96-well plates at a density of 8,000 cells/well and cultured at 37 ℃ for 24 h. Afer 24 h incubation, cells were treated with various concentrations of SDAP1 and SDAP2 (0.25, 0.5, 0.75, 1, 1.25 and 1 mg/mL) at 24 h, and aferwards exposed to GM (3 mg/mL) for 24 h. Next, afer 24 h of GM treatment, the MTT solution was added to each well, and incubated at 37 ℃ for 4 h. Te medium was removed, and 150 μL of dimethylsulfoxide (Solarbio, China) was added to each well. Finally, the optical density was detected with a microplate reader at 490 nm.
Reference：
Wang Z, Wang L, Wang J, Luo J, Ruan H, Zhang J. Purified Sika deer antler protein attenuates GM-induced nephrotoxicity by activating Nrf2 pathway and inhibiting NF-κB pathway. Sci Rep. 2020 Sep 24;10(1):15601. doi: 10.1038/s41598-020-71943-6. PMID: 32973191; PMCID: PMC7518274.
Cell（HepG2 Cell，5 mg/mL MTT，4h, 570nm）：
Methylthiazole tetrazolium (MTT) assay was applied to measure the cell viability of Up-3, Up-4, and Up-5. Briefly, after incubation, the cells was washed with PBS, and incubated with 10 μL of MTT (5 mg/mL) for 4 hours. The supernatants was mixed with 100 μL of DMSO and then the absorbance was measured at 570 nm with a microplate reader. The absorbance of the untreated cells (Normal group) was considered as 100%.
Reference：
Zhong QW, Zhou TS, Qiu WH, Wang YK, Xu QL, Ke SZ, Wang SJ, Jin WH, Chen JW, Zhang HW, Wei B, Wang H. Characterization and hypoglycemic effects of sulfated polysaccharides derived from brown seaweed Undaria pinnatifida. Food Chem. 2021 Mar 30;341(Pt 1):128148. doi: 10.1016/j.foodchem.2020.128148. Epub 2020 Sep 22. PMID: 33038776.
