Methyl thiazolyl tetrazolium (mtt)

Manufactured by Meilun

Sourced in China

MTT is a colorimetric assay used to measure the metabolic activity of cells. It is a widely used method for evaluating cell viability, proliferation, and cytotoxicity in various cell-based studies.

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Lab products found in correlation

Streptomycin, by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Trypsin, by Meilun (2 mentions) 96 well plate, by NEST Biotechnology (2 mentions) Sepharose cl 4b gel, by Solarbio (2 mentions) Crystalline violet, by Solarbio (1 mentions) Dextran sulfate sodium (dss), by Yeasen (1 mentions) 2 7 dichlorofluorescein diacetate dcfh da, by Beyotime (1 mentions) Epigallocatechin gallate (egcg), by Maclin Biochemical Technology (1 mentions) Curcumin, by Maclin Biochemical Technology (1 mentions) Cell culture medium, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Hoechst 33342, by BD (1 mentions) Propidium iodide, by BD (1 mentions) Annexin 5, by BD (1 mentions) Cm5 sensor chip, by GE Healthcare (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Coumarin 6, by Meilun (1 mentions) Sirna mate, by GenePharma (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Meilun (1 mentions)

12 protocols using methyl thiazolyl tetrazolium (mtt)

Cytotoxicity and Cell Cycle Assays

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MDCK-MDR1, MDCK-WT, and 4T1 were seeded in 96-well plates at a density of 8 × 10³/well for 24 h. The cell was treated with PTX (Meilunbio, Dalian, China, CAS 33069-62-4) and PIP (Meilunbio, CAS 94-62-2) dissolved in DMSO and diluted in the medium containing 1% FBS for 24 h and 48 h, respectively. Then, the medium was replaced with 200 μL 0.5 mg/mL MTT (Meilunbio, CAS 298-93-1) solution for 4 h. The MTT solution was carefully discarded and 100 μL DMSO was added to fully dissolve the crystals in a shaker at 37 °C. The inhibition rate of the drug to the cells was calculated by measuring the absorbance value at a wavelength of 490 nm.
4T1 cells were seeded in 12-well plates (5 × 10⁴/well) and treated with the drug for 24 h, then fixed with 70% ethanol. Cell cycle analysis was conducted with Propidium staining (HY-KY1071) and flow cytometry, and Crystalline Violet staining (Solarbio, Beijing, China, G1062), and via observation under a microscope.

Xu W., Xiao Y., Zheng L., Xu M., Jiang X, & Wang L. (2023). Enhancing Paclitaxel Efficacy with Piperine-Paclitaxel Albumin Nanoparticles in Multidrug-Resistant Triple-Negative Breast Cancer by Inhibiting P-Glycoprotein. Pharmaceutics, 15(12), 2703.

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Antioxidant and Anti-inflammatory Effects

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Active baker's Yeast was purchased from Angel Yeast Co., Ltd. (Yichang, China). Curcumin and EGCG were purchased from Macklin (Shanghai, China). DSS was purchased from Yeasen (Shanghai, China). Phosphate buffer saline (PBS) was purchased from Sevier (Wuhan, China). 2,7-dichlorofluorescein diacetate (DCFH-DA) was purchased from Beyotime Biotechnology Co., Ltd. Fluorescent lipophilic dyes (DiL and DiR) were purchased from Promokine (Heidelberg, Germany). MTT, myeloperoxidase (MPO) assay kit, lipopolysaccharides (LPS), and antibodies (CD86, CD80, and CD206) were purchased from Meilun Bio (Dalian, China).

Li J., Song J., Deng Z., Yang J., Wang X., Gao B., Zhu Y., Yang M., Long D., Luo X., Zhang M., Zhang M, & Li R. (2024). Robust reactive oxygen species modulator hitchhiking yeast microcapsules for colitis alleviation by trilogically intestinal microenvironment renovation. Bioactive Materials, 36, 203-220.

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Synthesis and Characterization of PPa-PEG2K Nanoparticles

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PPa was purchased in Shanghai Dibo Biotechnology Co., Ltd. Cell culture medium, penicillin, streptomycin and fetal bovine serum (FBS) were all obtained from GIBCO, Invitrogen Corp. (Carlsbad, CA, USA). MTT, trypsin, Singlet Oxygen Sensor Green Reagent (SOSG) and Reactive Oxygen Species Assay Kit (DCFH-DA) were purchased from Dalian Meilun Biological Technology Co., Ltd. Hoechst 33342 was purchased from BD Biosciences. Cell culture consumables were purchased by Wuxi NEST Biotechnology Co., Ltd. PCL-PEG_2K was obtained from Shanghai Ponsure Biological Technology Co., Ltd. PPa-PEG_2K was obtained by our previous synthetic method³¹ (link). Other reagents were of analytical grade.

Zhang S., Wang Y., Kong Z., Zhang X., Sun B., Yu H., Chen Q., Luo C., Sun J, & He Z. (2021). Pure photosensitizer-driven nanoassembly with core-matched PEGylation for imaging-guided photodynamic therapy. Acta Pharmaceutica Sinica. B, 11(11), 3636-3647.

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Paclitaxel-Loaded Nanoparticles Synthesis

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Paclitaxel (PTX) was purchased from Jingzhu Bio-technology Co. Ltd. (Nanjing, China). Succinic anhydride (SA), glutaric anhydride (GA), trans-2-butene-1,4-dicarboxylic acid (DA), N-(3-dimethylaminopropyl)-N-ethyl-carbodiimide hydrochloride (EDCI) and 4-dimethylaminopyrideine (DMAP) were provided by Aladdin Industrial Corporation (Shanghai, China). 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol (for injection) (Chol) and 2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methyl (polyethylene glycol)-2000 (DSPE-PEG2000) were purchased from Shanghai Advanced Vehicle Technology Pharmaceutical Ltd. (Shanghai, China). Sepharose CL-4B gel was purchased from Solarbio Corporation. Roswell Park Memorial Institute (RPMI-1640) and trypsin were obtained from Gibico (Beijing, China). Fetal bovine serum (FBS) was obtained from Foundation (Beijing, China), and MTT was purchased from Meilun Biotech Co. Ltd. (Dalian China). 96-well plates were supplied by NEST Biotechnology (Jiangsu, China). All other materials were of analytically pure grade.

Yu J., Zhou S., Li J., Wang Y., Su Y., Chi D., Wang J., Wang X., He Z., Lin G., Liu D, & Wang Y. (2020). Simple weak-acid derivatives of paclitaxel for remote loading into liposomes and improved therapeutic effects. RSC Advances, 10(46), 27676-27687.

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Apoptosis and Cell Cycle Assay

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AML cells were stained with Annexin-V and Propidiumiodide (PI) purchased from BD Biosciences and Cell Signaling Technology. Cell viability was determined by MTT (Meilunbio) and lysis solution (10% SDS, 5% isobutanol and 0.012 M HCL). Cells were fixed by 75% ice ethanol over 24 h before cell cycle assay, and then were stained with PI.

Hu C.L., Chen B.Y., Li Z., Yang T., Xu C.H., Yang R., Yu P.C., Zhao J., Liu T., Liu N., Shan B., Zhang Q., Song J., Fei M.Y., Zong L.J., Zhang J.Y., Wu J.C., Chen S.B., Wang Y., Chang B., Hou D., Liu P., Jiang Y., Li X., Chen X., Deng C.H., Ren Y.Y., Wang R., Jin J., Xue K., Zhang Y., Du M., Shi J., Wu L.Y., Chang C.K., Shen S., Chen Z., Chen S.J., Liu X., Sun X.J., Zheng M, & Wang L. (2022). Targeting UHRF1-SAP30-MXD4 axis for leukemia initiating cell eradication in myeloid leukemia. Cell Research, 32(12), 1105-1123.

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Evaluating Nanoxel-PM Delivery Efficacy

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DTX, ginsenoside 20 (S)–Rg3, and Nanoxel-PM (Samyang Biopharm) were gifted by Bensu Medicine Technology Co. Ltd. (Shanghai, China). Cholesterol was purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China). PL-100M was purchased from AVT Medicine Technology Co. Ltd. (Shanghai, China). Coumarin-6 (C6), Hoechst 3334w, DAPI, 1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide (DiR), DiD, MTT, and 1% crystal violet solution were purchased from Meilun Biotechnology Co., China. Annexin V–FITC/propidium iodide (PI) apoptosis detection kit and phosphatase inhibitor cocktail were provided by Yeasen Biotechnology Co., China. Penicillin, streptomycin, and 0.25% trypsin-EDTA were purchased from Invitrogen Co., USA. Protein GLUT1 was obtained from Wuhao Bio-Tech Co., China. CM5 sensor chips were purchased from GE Healthcare Bio-Sciences AB (Sweden). Anti-Glut1 small interfering RNA (siRNA) (CCAACUGGACCUCAAACUUTT, AAGUUUGAGGUCCAGU-UGGTT) and siRNA-mate were provided by GenePharma Co., China. All other chemicals and solvents used in this study were of reagent grade or high-performance liquid chromatography (HPLC) grade.

Xia J., Ma S., Zhu X., Chen C., Zhang R., Cao Z., Chen X., Zhang L., Zhu Y., Zhang S., Li S., Gu G., Wei X., Yu K, & Wang J. (2022). Versatile ginsenoside Rg3 liposomes inhibit tumor metastasis by capturing circulating tumor cells and destroying metastatic niches. Science Advances, 8(6), eabj1262.

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Combination Cancer Treatment Protocol

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Metformin, 2-deoxyglucose, doxorubicin, vera-pamil, digoxin, MTT were all purchased from Dalian Meilun Biotech Co., Ltd.(China). p53 activator AB143228 was purchased from Abcam Company. p53 inhibitor PFTα was obtained from Selleck Chemicals LLC.

Xue C., Wang C., Sun Y., Meng Q., Liu Z., Huo X., Sun P., Sun H., Ma X., Ma X., Peng J, & Liu K. (2016). Targeting P-glycoprotein function, p53 and energy metabolism: Combination of metformin and 2-deoxyglucose reverses the multidrug resistance of MCF-7/Dox cells to doxorubicin. Oncotarget, 8(5), 8622-8632.

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Doxorubicin-Loaded Nanoparticle Formulation

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Doxorubicin hydrochloride, MTT and DiR were supplied by Meilun Biotech Co. Ltd. (Dalian, China). 1,2-distearoyl-sn‑glycero-3-phosphoethanolamine-N-[amino(polyethyleneglycol)−2000] (DSPE-PEG2k), phosphatidylcholine (PC) and cholesterol were purchased from Shanghai Advanced Vehicle Technology Pharmaceutical Ltd., (Shanghai, China). Ferric ammonium citrate and ethylenediamine tetraacetic acid (EDTA) were obtained from Yuwang Pharm. (Shandong, China). GSH were offered by Aladdin (Shanghai, China). C11-BODIPY 581/591 were purchased from Maokang Biotechnology (Shanghai, China). ROS Assay Kit and GSH Assay Kit were offered by Beyotime (Shanghai, China). Sepharose CL-4B gel, hydroxyethyl piperazine ethylsulfonic acid (HEPES) and Hoechst 33,342 was provided by Solarbio Science and Technology Co. Ltd. (Beijing, China). 96-well plates were purchased from NEST Biotechnology (Wuxi, China). Fetal bovine serum (FBS) was obtained from Hyclone (Beijing, China).

Yang Y., Zuo S., Li L., Kuang X., Li J., Sun B., Wang S., He Z, & Sun J. (2021). Iron-doxorubicin prodrug loaded liposome nanogenerator programs multimodal ferroptosis for efficient cancer therapy. Asian Journal of Pharmaceutical Sciences, 16(6), 784-793.

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Antiproliferative Activity of Compounds

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The in vitro antiproliferative activity of the target compounds and CA-4 was measured by a standard MTT (meilunbio®, China) assay. Cervical cancer (HeLa), gastric adenocarcinoma (SGC-7901) and breast cancer (MCF-7) were used, respectively. Cells were inoculated in 96-well plates at a density of 2 × 10³/well. After 24 h, the target concentration of drug was added to each well and incubated for 72 h at 37 °C under 5% CO₂. 20 μL of fresh medium containing 5 mg/ml MTT solution was added and incubation continued for 4 h. After removing the medium containing MTT from each well, 150 μL of dimethyl sulfoxide (DMSO) was added to each well until the purple formazan crystals was completely dissolved, placed in a multimode plate reader Victor Nivo 3S (Perkinelmer, USA) and the absorbance was measured at 490 nm.²⁶ (link)

Yang S., Wang C., Shi L., Chang J., Zhang Y., Meng J., Liu W., Zeng J., Zhang R., Shao Y, & Xing D. (2022). Design, synthesis and biological evaluation of novel diarylpyridine derivatives as tubulin polymerisation inhibitors. Journal of Enzyme Inhibition and Medicinal Chemistry, 37(1), 2755-2764.

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Hydrogel-Based Biosensing Platform

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Sodium hyaluronate (HA, average molecular mass of 100 to 200 kDa), GOx, HRP, pyrrole, DA, RhB, H&E staining solution, and LB broth were purchased from Shanghai Macklin Biochemical Co. Ltd. Zinc nitrate hexahydrate [Zn(NO₃)₂·6H₂O] and 2-methylimidazole were acquired from Aladdin. MTT, living/dead cell double-staining kit (calcein-AM/PI), and BCA protein assay kit were obtained from Dalian Meilun Biotechnology Co.

Zhang X., Wang Z., Jiang H., Zeng H., An N., Liu B., Sun L, & Fan Z. (2023). Self-powered enzyme-linked microneedle patch for scar-prevention healing of diabetic wounds. Science Advances, 9(28), eadh1415.

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