3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bro- mide (MTT) reagent (MP Bio, Santa Ana, CA, USA) dissolved in 1X PBS (5 mg/mL) was added to culture medium to a final concentration of 0.16 mg/mL. Cells were incubated for 4 h at 37°C, after which media and MTT reagent were removed. Formazan crystals were solubilized in 1 mL DMSO, diluted 1:10 in additional DMSO, then transferred to a microplate to quantify A570 using a Biotek plate reader.
Mtt reagent
Manufactured by MP Biomedicals
Sourced in United States
The MTT reagent is a colorimetric assay used to measure the metabolic activity of cells. It is a versatile tool for various applications in cell biology and biochemistry research.
Automatically generated - may contain errors
Lab products found in correlation
Fluostar omega, by BMG Labtech (1 mentions)
Glycine, by MP Biomedicals (1 mentions)
Lc ms 6410, by Agilent Technologies (1 mentions)
Tris base, by MP Biomedicals (1 mentions)
Acrylamide, by MP Biomedicals (1 mentions)
Ponceau s, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by MP Biomedicals (1 mentions)
Fluostar omega plate reader, by BMG Labtech (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by MP Biomedicals (1 mentions)
Elx800, by Agilent Technologies (1 mentions)
8 protocols using mtt reagent
1
MTT Cell Viability Assay Protocol
2
MTT Assay for Cell Viability
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MTT assay was performed as described earlier [60 (link),61 (link),71 (link)]. Briefly, cells (5000 cells/well) were seeded in triplicates in 96-well plates and treated with various concentrations of SP11 after a 24 h incubation. Cells were then treated with MTT reagent, MP Biomedicals (5 mg/mL) at 37 °C, 5% CO2, post 48 and 72 h of incubation with SP11. Absorbance was recorded at 570 nm and the results shown were collected from three different biological replicates.
3
Combinatorial Effects of Cisplatin and ST08 on Cell Viability
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MTT assay was performed as described earlier (2 (link), 5 (link)). Cells were seeded (5000 cells/well) in a 96-well plate in triplicates, incubated for 24 h, and treated with various concentrations of Cisplatin (0,1,5,10,20,50 µM) and ST08 (30,60nM). Cells were incubated with MTT reagent, MP Biomedicals (5 mg/ml) after 48h of incubation of cells with the drugs at 37°C and 5% CO2. Absorbance was measured at 570 nm, and the results shown are from 3 different biological replicates. To understand the effect of the combination of Cisplatin with ST08, the combination index method (38 (link), 39 (link)) was used. The combination index is calculated as follows:
where CAX, and CBX are the concentration of drugs A and B used in combination to achieve x % drugeffect. ICX,A and ICX,B are the concentrations for single agents to achieve the same effect. CI < 1 implies Synergism, CI = 1 is Additive and CI > 1 is Antagonistic effect.
where CAX, and CBX are the concentration of drugs A and B used in combination to achieve x % drugeffect. ICX,A and ICX,B are the concentrations for single agents to achieve the same effect. CI < 1 implies Synergism, CI = 1 is Additive and CI > 1 is Antagonistic effect.
4
Cell Proliferation and Viability Assay
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Cell survival/proliferation was performed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent (MP Biochemicals, Santa Ana, CA). Cells (2–5 × 105) were seeded onto 6-well plates overnight and then transfected with 50 nM NC or miR-214 mimic (or 2 µg EV or PTK6) for 24 h. The transfected cells were then harvested and seeded onto 96-well plates (2.5–5 × 103 cells/well) and grown for 24, 48, or 72 h. For ibrutinib (IBT) experiments, transfected cells were seeded onto 96-well plates and treated with the indicated concentrations of IBT for 48 h. Following exposure to specific transfection/treatments, cells were incubated with 10 µl/well of MTT reagent (5 mg/ml) for 2 h at 37 °C in a cell culture incubator. Formazan crystals were dissolved in dimethyl sulfoxide (DMSO), and cell survival/proliferation was quantified by reading the plates at 570 nm using a Fluostar Omega plate reader (BMG Lab Tech, Cary, NC).
5
Cytotoxicity Evaluation of ST09
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The MTT assay was performed as described earlier [13 (link)]. Cells (5000 cells/well) were seeded in triplicates in 96-well plates. After a 24 h incubation, cells were treated with various concentrations of ST09. Cells were treated with MTT reagent, MP Biomedicals (5 mg/mL) at 37 °C, 5% CO2, after 24, 48, and 72 h of incubation with ST09 as described in [13 (link)]. Absorbance was recorded at 570 nm and the results shown were collected from three different biological replicates.
6
Spectroscopic Characterization of 3-(2-Bromoacetyl)-2H-Chromen-2-One
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The IR spectra were recorded in KBr on a Jasco 430+; the 1H NMR spectra were recorded in CDCl3/DMSO on a Bruker (400 or 100 MHz) and J values were reported in hertz (Hz) for 1H NMR. 3-(2-bromoacetyl)-2H-chromen-2-one was prepared as per the literature [77 (link)]. The LCMS spectrum of SP11 was recorded on triple-quadrupole liquid chromatography mass spectrometry (LC-MS) 6410 from Agilent Technologies.
Ponceau S (Sigma life Sciences, Burlington, MA, USA), Phenylmethanesulfonyl chloride (PMSF), protease inhibitor cocktail tablets (EDTA-free), Tris base, Glycine, acrylamide, and bis-acrylamide powder, DMSO, MTT reagent and all other routine chemicals were purchased from MP biomedicals, (Santa Ana, CA, USA).
Ponceau S (Sigma life Sciences, Burlington, MA, USA), Phenylmethanesulfonyl chloride (PMSF), protease inhibitor cocktail tablets (EDTA-free), Tris base, Glycine, acrylamide, and bis-acrylamide powder, DMSO, MTT reagent and all other routine chemicals were purchased from MP biomedicals, (Santa Ana, CA, USA).
7
Evaluating TNFα-induced cytotoxicity in skin cancer cells
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Skin cancer A431, A375, A2058 and, normal skin HaCaT cells (1 × 104 cells/well) were grown in 96 plates for 18 h. Cells were treated with different concentrations of TNFα for 48 h and relative cell survival was analyzed by MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent (MP Biochemicals, Santa Ana, CA). Similarly, in other experiments, skin cancer cells or normal skin cells (1 × 105) were seeded onto 6-well plates for 24 h and transfected with EV of TNFAIP8-Myc plasmids or control siRNA and TNFAIP8 siRNA separately for 24 h. The transfected cells were harvested, and ten thousand cells seeded onto 96-well plates 48 h. Cells were incubated with 5 µl/well of MTT reagent (5 mg/ml) for 1 h at 37 °C in a cell culture incubator. Cells were washed with PBS, formazan crystals were dissolved in DMSO, and cell survival was quantified by reading the plates at 570 nm using a Fluostar Omega plate reader (BMG Lab Tech, Cary, NC).
8
Cytotoxicity and Proliferation Assays of MCL Cells
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