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MTT是一种广泛用于测量细胞增殖的比色剂。MTT可以被线粒体内的一些脱氢酶还原生成结晶状的深紫色产物formazan，可以被DMSO完全溶解，然后通过酶标仪可以测定490nm波长附近的吸光度。细胞增殖越多越快，则吸光度越高；细胞毒性越大，则吸光度越低。

注意事项：MTT溶液需避光保存，长时间光照会导致失效。当颜色变为灰绿色时，请勿使用。

使用说明（仅供参考）

1.收集对数期细胞，根据自己的实验需求，培养细胞。
2.小心吸去上清，加入90μL新鲜培养基，再加入10μL MTT溶液，继续培养4 h。
3.然后吸掉上清，每孔加入110μL DMSO，置摇床上低速振荡10 min，使结晶物充分溶解。

4.在酶联免疫检测仪490 nm处测量各孔的吸光值。

注：MTT溶液浓度一般的常用浓度为0.5%（仅供参考），实验者也可根据自己的实验需求进行调整。


使用本产品的应用案例（仅供参考）

In Vitro

Cell （HEK-293 cells，37 °C ，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 °C 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 °C 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.

来源文献：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%.

来源文献：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.

Cell ( A549 or A549/DDP cells，20 μL MTT (5 mg·mL⁻¹) was added to the wells for 4 h in an incubator )

The MTT assay was used to evaluate in vitro cytotoxicity of the drugs. Briefly, A549 or A549/DDP cells were seeded into 96-well plates (5,000 cells per well) and treated with various concentrations of the drugs for 48 h. Then, 20 μL MTT (5 mg·mL⁻¹) was added to the wells for 4 h in an incubator. The formazan formed from MTT was dissolved by adding 150 μL DMSO. 

来源文献：Ye W, Lv H, Zhang Q, Zhao J, Zhao X, Zhao G, Yan C, Sun F, Zhao Z, Jia X. A cisplatin and disulphiram co-loaded inclusion complex overcomes drug resistance by inhibiting cancer cell stemness in non-small cell lung cancer. J Drug Target. 2024 Dec;32(2):159-171. doi: 10.1080/1061186X.2023.2298844. Epub 2024 Feb 1. PMID: 38133515.

Cell（The RAW 264.7 cells；1% MTT incubated in the dark for 4 h）

Following a 24 h incubation period at 37˚C, a 1% MTT solution was added to each well and cells were incubated in the dark for 4 h. Thereafter, the medium was aspirated and replaced with 100 µl of DMSO in each well. Following a 10 min incubation at 37˚C, the absorbance was determined at a wavelength of 570 nm using a

microplate reader .

来源文献：Cheng X, Sun Q, Zheng R, Sun Y, Liang Z, Li N, Deng C. Milk-derived exosomes exert anti-inflammatory activity in lipopolysaccharide-induced RAW264.7 cells by modulating the TLR4/NF-κB and PI3K/AKT signaling pathways. Exp Ther Med. 2025 May 29;30(2):149. doi: 10.3892/etm.2025.12899. PMID: 40502622; PMCID: PMC12150682.

Cell（3 T3 and RAW264.7 cells，0.5 mg/mL MTT solution，4 h）

3 T3 and RAW264.7 cells were seeded in 96-well plates at 5 × 10³ cells/well and incubated overnight in RPMI-1640 medium. The cells were then exposed to various concentrations of DWN nanozymes (0, 6.25, 12.5, 25, 50, 100, and 200 μg/mL) for 24 h. After incubation, MTT solution (0.5 mg/mL) was added to each well and incubated for another 4 h. Then, 100 μL of DMSO was added, and the absorbance at 490 nm was recorded using a microplate reader.

文献来源：Liu R, Wu Y, Qu H, et al. Magnetically and optically powered nanozymes: Boosting breast cancer therapy with magnetothermodynamic-enhanced ferroptosis and apoptosis[J]. Chemical Engineering Journal, 2025, 509: 161353.

Cell（ID8 cells；10%；37℃；490 nm）

The cytotoxicity of the drugs was assessed using MTT method. Twelve hours later, the supernatant was discarded, 10% MTT (100 μL well−1) solution was added, and incubated for 4 h in a light-avoidance incubator at 37℃, and DMSO was used to dissolve the generated formazan. Finally, optical density (OD) value of each group at 490 nm was detected by enzyme labeller.

文献来源：Fu H, Liu X, Fang F, Pan Z, Chen L, Chen G, Yang X, Wang W, Yang B, Chen J, Yang X, Yu Z, Liang D, Yuan S. Amplification of cGAS-STING pathway with "single-molecule multitarget" nanoparticles for chemo-immunotherapy of ovarian cancer. Biomaterials. 2025 Dec;323:123434. doi: 10.1016/j.biomaterials.2025.123434. Epub 2025 May 24. PMID: 40446710.

In Vivo

Mice(Female Balb/c,6–8 weeks, subcutaneous injection,20 μL of MTT solution (5 mg/mL) for 4 h)

4T1 cells were seeded onto 96-well plates at 8000 cells per well. After 24 h of cultivation, the media was removed. 0.2 mL of PTX injections, DP NPs, or DPB NPs, diluted with the incomplete medium, were added to the 96-well plates. The blank incomplete medium was the negative control.

After 72h co-incubation, each well was treated with 20 μL of MTT solution (5 mg/mL) for 4 h. After removing the medium, 0.2 mL DMSO was added to each well. The absorbance of each well was measured at 570 nm using a microplate reader.

文献来源：Li M, Chen M, Li P, Zhang Z, Yu H, Wang X. Enhanced transcytosis and therapeutic efficacy of paclitaxel nanoparticles: Pyridylboronic acid modification and sialic acid targeting. Colloids Surf B Biointerfaces. 2024 Nov 30;247:114417. doi: 10.1016/j.colsurfb.2024.114417. Epub ahead of print. PMID: 39631321.

Fu H,et al.Biomaterials. 2025 Dec;323:123434. doi: 10.1016/j.biomaterials.2025.123434.

Zhong QW, et al. Food Chem. 2021 Mar 30;341(Pt 1):128148.