Infinite m2000 pro plate reader

Manufactured by Tecan

Sourced in Germany

The Infinite M2000 Pro is a multi-mode microplate reader from Tecan. It is designed to accurately measure various assays in microplate format, including absorbance, fluorescence, and luminescence detection. The device features a high-performance monochromator-based optical system and can accommodate a wide range of microplate types and sample volumes.

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Lab products found in correlation

Resazurin, by Merck Group (5 mentions) Resazurin, by Promega (4 mentions) Resazurin, by Tecan (3 mentions) Prism 5, by GraphPad (2 mentions) Trypsin edta, by Thermo Fisher Scientific (2 mentions) 96 well cell culture plate, by Thermo Fisher Scientific (2 mentions) Bromocriptine, by Merck Group (1 mentions) Triptolide, by Merck Group (1 mentions) Elisa based kit, by Active Motif (1 mentions) Sensolyte sars cov 2 3cl protease activity assay kit, by AnaSpec (1 mentions) Resorufin, by Tecan (1 mentions) Dual glo luciferase reporter assay system, by Promega (1 mentions) Cignal myc reporter assay kit, by Qiagen (1 mentions) Niclosamide, by Merck Group (1 mentions) Rat tail collagen 1, by Thermo Fisher Scientific (1 mentions) Prism v6, by GraphPad (1 mentions) 2 7 dichlorodihydrofluorescein diacetate h2dcf da, by Merck Group (1 mentions) Tween 20, by Merck Group (1 mentions) Crizotinib, by Merck Group (1 mentions) Resazurin solution, by Merck Group (1 mentions)

Close spelling variants of this product

Infinite M2000 Pro (42 citations) Infinite plate reader (36 citations) Infinite M2000 (25 citations) Infinite M2000 ProTM plate reader (8 citations) Infinite M2000 ProTM (4 citations) M2000 (3 citations) Infinite M2000 Pro plate (2 citations)

25 protocols using infinite m2000 pro plate reader

Cytotoxicity Assay of Bromocriptine Combination

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The cytotoxic effects of bromocriptine (Sigma-Aldrich, Taufkirchen, Germany; Figure 7A) was evaluated by the resazurin assay (O’Brien et al., 2000 (link)). This assay is based on reduction of the indicator dye, resazurin, to the highly fluorescent resorufin by viable cells. Aliquots of 5,000 cells/100 μL of HEK293 and MDA-MB-231 were seeded in 96-well plates and incubated for one day before treatment. However, for leukemic cells, 10,000 cells/100 μL cells placed into 96-well plates and immediately treated. After 72 h incubation, 20 μL resazurin 0.01% w/v solution were added to each well, and the plates were incubated at 37°C for 4 h. Fluorescence was measured by an Infinite M2000 Proplate reader (Tecan, Crailsheim, Germany) using an excitation wavelength of 544 nm and an emission wavelength of 590 nm. Each experiment was done at least three times with six replicates each. The viability was analyzed based on a comparison with untreated cells. Fifty percent inhibition (IC₅₀) values indicate the drug concentrations required to inhibit 50% of cell proliferation and were calculated from a calibration curve by linear regression using Microsoft Excel (Kuete et al., 2016a (link),b (link)).
A combination of 20 or 40% of the IC₅₀ value of bromocriptine (2.4 and 4.8 μM) with different concentrations of doxorubicin or paclitaxel was used to treat CEM/ADR5000 cells.

Seo E.J., Sugimoto Y., Greten H.J, & Efferth T. (2018). Repurposing of Bromocriptine for Cancer Therapy. Frontiers in Pharmacology, 9, 1030.

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Evaluating Triptolide's Cytotoxic Effects on Cancer Cells

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The cytotoxic activities of triptolide (Sigma Aldrich, Taufkirchen, Germany; Fig. 1A) were evaluated by the resazurin assay [25 (link)]. This assay is based on reduction of the indicator dye, resazurin, to the highly fluorescent resorufin by viable cells. Aliquots of 5,000 cells/100 µL of U87.MG and U87.MGΔEGFR were placed in 96-well plates and incubated for one day before treatment. However, for leukemic cells, 10,000 cells/100 µL cells were seeded into 96-well plates and immediately treated. Twenty microliters of resazurin 0.01% w/v solution were added to each well after 72 h at 37 °C incubation, and the plates were incubated at 37 °C for 4 h. Fluorescence was detected by an Infinite M2000 Proplate reader (Tecan, Crailsheim, Germany) with an excitation wavelength of 544 nm and an emission wavelength of 590 nm. Each experiment was carried out at least three times with six replicates each. The viability was analyzed based on a comparison with untreated cells. Fifty percent inhibition (IC₅₀) values imply the drug concentrations needed to inhibit 50% of cell proliferation and were calculated from a calibration curve by linear regression using Microsoft Excel [26 , 27 ].Fig. 1

Cytotoxicity of triptolide against cancer cells. Chemical structure of triptolide A. Cytotoxic effect of triptolide against CEM/ADR5000 and CCRF-CEM B, and U87.MG and U87.MGΔEGFR C

Seo E.J., Dawood M., Hult A.K., Olsson M.L, & Efferth T. (2021). Network pharmacology of triptolide in cancer cells: implications for transcription factor binding. Investigational New Drugs, 39(6), 1523-1537.

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Influenza Virus Infection Assay

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The assay was performed as described by Noah et al. [38 (link)] with some modifications. MDCK cells were seeded into 96-well plates, incubated overnight and infected with influenza virus (MOI = 0.1) suspended in DMEM supplemented with 1% FBS, test compound and 2 μg/mL TPCK-treated trypsin, with a final DMSO concentration of 1% in each well, followed by 40 h of incubation, CellTiter-Glo reagent was added, and the plates were read using a Tecan Infinite M2000 PRO™ plate reader.

Liang S., Li M., Yu X., Jin H., Zhang Y., Zhang L., Zhou D, & Xiao S. (2019). Synthesis and structure-activity relationship studies of water-soluble β-cyclodextrin-glycyrrhetinic acid conjugates as potential anti-influenza virus agents. European Journal of Medicinal Chemistry, 166, 328-338.

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Resazurin-Based Cell Viability Assay

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Cell viability was measured using the resazurin assay as previously described in [72 (link)]. Human diploid MRC-5 lung fibroblasts was kindly provided by Dr. rer. nat. Sebastian Zahnreich (Department of Radiation Oncology and Radiation Therapy, University Medical Center of the Johannes Gutenberg University, Mainz, Germany) were seeded (5 × 10⁵ cells per well) into 96-well culture plates and incubated overnight before treatment. On the second day, the cells were treated with 10 concentrations of the four compounds in a range of 0.3–100 μM. After 72 h incubation, 20 μL 0.01% resazurin (Promega, Mannheim, Germany) was added to each well. Fluorescence was detected after 4 h incubation using an Infinite M2000 Pro plate reader (Tecan) at Ex/Em = 550 nm/590 nm wavelength. Cell viability was calculated in comparison to DMSO control. The DMSO final concentration was 0.5%. The 50% cytotoxicity concentration (CC₅₀) values were calculated in comparison to the DMSO-treated control. Each experiment was independently repeated three times with six wells for each concentration. Therapeutic indices were calculated using the following equation: Therapeutic index = TD₅₀/ED₅₀.

Shahhamzehei N., Abdelfatah S, & Efferth T. (2022). In Silico and In Vitro Identification of Pan-Coronaviral Main Protease Inhibitors from a Large Natural Product Library. Pharmaceuticals, 15(3), 308.

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Resazurin Assay for Cytotoxicity Assessment

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The non-fluorescent dye resazurin is reduced metabolically by living cells to the strongly-fluorescent dye resorufin (O'Brien et al., 2000 (link)). The resazurin (Promega, Mannheim, Germany) reduction assay was performed to assess the cytotoxicity of sanguinarine toward drug-sensitive and -resistant cell lines. Briefly, tumor cells (2 × 10⁴cells/well) were seeded in 96-wells plate in a volume of 100 μL, and varying concentrations of sanguinarine were added to reach the total volume of 200 μL. After 72 h, 20 μL of 0.01% w/v resazurin (Sigma-Aldrich, Schnelldorf, Germany) was added to each well. Cells were incubated for 4 h at 37°C. Fluorescence at excitation wave length 544 nm and emission at 590 nm was measured using Infinite M2000 Pro™ plate reader (Tecan, Crailsheim, Germany). The percentage of viable cells was calculated as follows:

\begin{array}{l} Cell Viability (% of control) = \\ \frac{a b s o r p t i o n (t r e a t e d c e l l s) - a b s o r p t i o n (m e d i u m a l o n e)}{a b s o r p t i o n (u n t r e a t e d c e l l s) - a b s o r p t i o n (m e d i u m a l o n e)} * 100 \end{array}

IC₅₀ values were calculated from concentration dependent curves using nonlinear regression analysis tool built in Prism 7 GraphPad software. All IC₅₀ values are expressed as the mean ± standard deviation (SD). Each assay was repeated thrice independently with six replicates each.

Saeed M.E., Mahmoud N., Sugimoto Y., Efferth T, & Abdel-Aziz H. (2018). Molecular Determinants of Sensitivity or Resistance of Cancer Cells Toward Sanguinarine. Frontiers in Pharmacology, 9, 136.

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c-MYC DNA Binding Activity Assay

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The c-MYC DNA binding activity assays were performed using TransAM enzyme-linked Immunosorbent assay (ELISA)-based kits (Active Motif, Rixensart, Belgium) according to the manufacturer's protocol. Briefly, 10 μg of nuclear extracts from control, shikonin, derivatives, 10074-G5 or 10058-F4-treated cells were separately incubated in a 96-well plate immobilized with an oligonucleotide containing the c-MYC consensus binding site (5′-CACGTG-3′). The active forms of transcription factors from extracts, which specifically bound to this oligonucleotide, were detected by a primary antibody against c-MYC in an ELISA-like format. The absorbance of the sensitive colorimetric reaction mediated by a secondary HRP-conjugated antibody was measured on the Infinite M2000 Proplate reader (Tecan) at 450 nm with a reference wavelength of 655 nm.

Zhao Q., Assimopoulou A.N., Klauck S.M., Damianakos H., Chinou I., Kretschmer N., Rios J.L., Papageorgiou V.P., Bauer R, & Efferth T. (2015). Inhibition of c-MYC with involvement of ERK/JNK/MAPK and AKT pathways as a novel mechanism for shikonin and its derivatives in killing leukemia cells. Oncotarget, 6(36), 38934-38951.

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Cytotoxicity Evaluation of PT Compound

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The cytotoxicity of PT was evaluated using the resazurin (Promega, Mannheim, Germany) reduction assay as previously described (Kuete et al., 2016 (link), 2017 (link)). Only viable cells can reduce and convert resazurin to highly fluorescent resorufin, while dead cells cannot convert resazurin dye (O’brien et al., 2000 (link)). Based on this principle, tumor cells were treated with different concentrations of PT and incubated for 72 h. An Infinite M2000 Proplate reader (Tecan, Germany) was used to measure the fluorescence using excitation/emission wavelength of 544/590 nm. The 50% inhibition concentrations (IC₅₀) were determined using dose response curves of each cell lines using Excel 2013 software (Microsoft, Redmond, WA, United States). The experiments were conducted three times independently with six replicates each.
The tumor cell line panel of the National Cancer Institute (NCI, United States) was treated with PT and subjected to the sulforhodamine B assay (Rubinstein et al., 1990 (link)).

Dawood M., Ooko E, & Efferth T. (2019). Collateral Sensitivity of Parthenolide via NF-κB and HIF-α Inhibition and Epigenetic Changes in Drug-Resistant Cancer Cell Lines. Frontiers in Pharmacology, 10, 542.

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Screening SARS-CoV Protease Inhibitors

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Enzymatic assays were performed using the SensoLyte SARS-CoV-2 3CL Protease Activity Assay Kit (AnaSpec, San Francisco, CA, USA), SARS-CoV-1 Assay Kit, and 3CL Protease MERS-CoV Assay Kit (BPS Bioscience, San Diego, CA, USA). Twelve selected compounds were diluted in assay buffer to a final concentration of 1 mM. Compound aliquots of 10 μL were added to black 96-well plates (Greiner, Frickenhausen, Germany), and 40 μL of 0.1 mg/mL M^pro was added to each well of the plates and incubated with the compounds at 37 °C for 30 min. The enzymatic reactions were initiated by adding a fluorescent substrate. The final concentration of the compound was 100 µM. Fluorescence was measured using an Infinite M2000 Pro plate reader (Tecan, Crailsheim, Germany). All values were subtracted from blank values. Then, compounds exhibiting more than 50% inhibitory activity at a fixed concentration of 100 µM were selected for dose-response studies in a concentration range from 0 to 100 µM for SARS-CoV-2, SARS-CoV-1, and MERS-CoV to calculate 50% inhibition concentrations (IC₅₀). The activity percentage of M^pro was calculated using the following equation: Activity % = 100 − [(RFU_{Vehicle control} − RFU_{tested sample})/RFU_{Vehicle control} × 100].

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Cytotoxicity Assay of Natural Compounds

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The cytotoxicity assay performed using resazurin reduction assay was applied to the crude extract (DCB), compounds 1-4, and doxorubicin [18 (link), 20 (link), 21 (link)] with similar experimental conditions as those reported earlier [13 (link), 19 (link), 22 (link), 23 (link)]. The Infinite M2000 Pro™ plate reader (Tecan, Crailsheim, Germany) with excitation wavelength of 544 nm and an emission wavelength of 590 nm was used to read the fluorescence after 72 h incubation. IC₅₀ values earlier defined [13 (link)] were calculated from a calibration curve by linear regression using Microsoft Excel [24 (link)]. The degree of resistance (D.R.) was determined as the IC₅₀ value of the resistant cell line versus that of its sensitive congeners; meanwhile, the selectivity index (S.I.) was the IC₅₀ value in normal AML12 hepatocytes versus that in HepG2 hepatocarcinoma.

Mbaveng A.T., Damen F., Simo Mpetga J.D., Awouafack M.D., Tane P., Kuete V, & Efferth T. (2019). Cytotoxicity of Crude Extract and Isolated Constituents of the Dichrostachys cinerea Bark towards Multifactorial Drug-Resistant Cancer Cells. Evidence-based Complementary and Alternative Medicine : eCAM, 2019, 8450158.

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Adapalene Cytotoxicity in MM, Leukemia, and PBMCs

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The sensitivity of MM cells, leukemia cells, and PBMCs to adapalene was evaluated by the resazurin reduction assay, as previously described [30 (link)]. In brief, 10⁴ cells were seeded in each well of a flat bottom 96-well plate. Cells were directly treated with 10 different concentrations of adapalene (Activate Scientific, Prien am Chiemsee, Bavaria) which are 3-fold apart from one another, ranging from 100 µM to 0.003 µM for MM and T-ALL cells. However, when 10-fold apart from one another, they ranged from 100 µM to 10⁻⁷ µM for PBMCs. The plates were incubated for 72 h at 5% CO₂/37 °C and then re-incubated for another 4 h under the same conditions, with 20 μL of resazurin (0.01% w/v; Sigma-Aldrich) added to each well. An Infinite M2000 Pro plate reader (Tecan, Crailsheim, Germany) was used to measure resorufin fluorescence, produced by the reduction of resazurin by live cells, at 544–590 nm (excitation-emission wavelengths). Afterward, cell viability was plotted vs. adapalene concentration, and the IC₅₀ values from three separate experiments with six repeats each were calculated with GraphPad Prism 5 software (GraphPad Software, San Diego, CA, USA).

Boulos J.C., Chatterjee M., Shan L, & Efferth T. (2023). In Silico, In Vitro, and In Vivo Investigations on Adapalene as Repurposed Third Generation Retinoid against Multiple Myeloma and Leukemia. Cancers, 15(16), 4136.

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