MTT 噻唑蓝

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.