MTT是一種廣泛用于測量細(xì)胞增殖的比色劑。MTT可以被線粒體內(nèi)的一些脫氫酶還原生成結(jié)晶狀的深紫色產(chǎn)物formazan，可以被DMSO完全溶解，然后通過酶標(biāo)儀可以測定490nm波長附近的吸光度。細(xì)胞增殖越多越快，則吸光度越高；細(xì)胞毒性越大，則吸光度越低。

注意事項：MTT溶液需避光保存，長時間光照會導(dǎo)致失效。當(dāng)顏色變?yōu)榛揖G色時，請勿使用。

使用說明（僅供參考）

1.收集對數(shù)期細(xì)胞，根據(jù)自己的實驗需求，培養(yǎng)細(xì)胞。
2.小心吸去上清，加入90μL新鮮培養(yǎng)基，再加入10μL MTT溶液，繼續(xù)培養(yǎng)4 h。
3.然后吸掉上清，每孔加入110μL DMSO，置搖床上低速振蕩10 min，使結(jié)晶物充分溶解。

4.在酶聯(lián)免疫檢測儀490 nm處測量各孔的吸光值。

注：MTT溶液濃度一般的常用濃度為0.5%（僅供參考），實驗者也可根據(jù)自己的實驗需求進行調(diào)整。


使用本產(chǎn)品的應(yīng)用案例（僅供參考）

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.

來源文獻：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%.

來源文獻：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.