Genios reader

Manufactured by Tecan

Sourced in Switzerland

The GENios reader is a compact and versatile microplate reader designed for a wide range of spectrophotometric applications in life science research. It can perform absorbance measurements across multiple wavelengths to support various assays and experiments.

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Lab products found in correlation

Glutathione reductase, by Merck Group (1 mentions) Bca protein quantification kit, by Interchim (1 mentions) Glutathione (gsh), by Merck Group (1 mentions) Erastin, by Merck Group (1 mentions) Celltiter blue, by Promega (1 mentions) Mtt labeling agent, by Merck Group (1 mentions) Celltiter 96 aqueous one solution, by Promega (1 mentions) Glucose assay kit, by Merck Group (1 mentions) Celltiter glo luminescent cell viability assay kit, by Promega (1 mentions) Dy501, by R&D Systems (1 mentions) Transparent 96 well plate, by Thermo Fisher Scientific (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Cell proliferation kit 2, by Roche (1 mentions) Cell proliferation kit 1, by Roche (1 mentions)

13 protocols using genios reader

Cellular ATP and Glucose Metabolism Assays

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Cellular ATP levels were determined using a CellTiter-Glo^® luminescent cell viability assay kit (Promega). Briefly, 4 × 10³ HeLa cells were seeded on an opaque-walled 96-well microplate and transfected for 120 h at 37°C with non-targeting or MMAB-targeting siRNA. After equilibrating microplates via incubation for 30 min at RT, 100-µl CellTiter-Glo^® reagent was added to 100-µl medium containing cells, and the plate was rotated for two minutes on an orbital shaker to induce cell lysis. Finally, plates were incubated for ten minutes at RT to stabilize the luminescent signal. The cells were imaged and recorded using a GENios reader (Tecan, Switzerland).
A glucose assay kit (Sigma-Aldrich) was used to measure glucose consumption. Briefly, 1 × 10⁵MMAB siRNA-transfected HeLa cells in a 6-well plate were incubated for 60 h at 37°C. Then, the medium was mixed with glucose assay reagent, transferred into 96-well microplates, and incubated for 30 min at 37°C. Microplates were measured at 540-nm using a GENios reader (Tecan).

Song K., Lee H.S., Jia L., Chelakkot C., Rajasekaran N, & Shin Y.K. (2022). SMAD4 Controls Cancer Cell Metabolism by Regulating Methylmalonic Aciduria Cobalamin Deficiency (cbl) B Type. Molecules and Cells, 45(6), 413-424.

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Quantification of Cellular Glutathione Levels

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Following two washing steps with ice-cold PBS, 2 × 10⁵ cells were scraped off in 200 µL ice-cold PBS containing 2 mM EDTA. Cell suspensions were lysed with 3% sulfosalicylic acid for 10 min on ice and centrifuged for 10 min at 14,000× g and 4 °C. The supernatant was incubated with triethanolamine/H₂O (1:1) and loaded with assay mix (300 µM DTNB, 450 µM NADPH, 20 units glutathione reductase (Sigma)) on a 96-well plate. A standard curve was generated by measuring different concentrations of pure GSH (Sigma). Absorption was measured at 390 nm for 26 min with a Tecan GENios reader. To determine total protein concentration, pellets were solved in 0.2 N NaOH and incubated overnight at 37 °C and subsequently measured using the bicinchoninic acid (BCA) protein quantification kit (Interchim, Montluçon, France) and BSA as standard. Statistical significance was calculated with a paired t-test (two-tailed) using GraphPad Prism software (n = 5; * p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001).

Nguyen L., Thewes L., Westerhoff M., Wruck W., Reichert A.S., Berndt C, & Adjaye J. (2023). JNK Signalling Regulates Self-Renewal of Proliferative Urine-Derived Renal Progenitor Cells via Inhibition of Ferroptosis. Cells, 12(17), 2197.

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Erastin-Induced Ferroptosis Modulation

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First, 1 × 10⁴ cells per well were seeded in a 96-well plate and treated with 0.1–100 µM erastin (Sigma-Aldrich) at 37 °C and 5% CO₂. After an incubation of 24 h, CellTiter-Blue^® (CTB, Promega, Madison, WI, USA) reagent was added to the cells at a ratio of 1:6 with fresh medium and incubated at 37 °C for 1 h. erastin-treated cells were additionally induced with 10 µM AEG3482 and the combination of 10 µM AEG3482 with 100 nM Lip-1. The change in resaruzin absorbance was measured with a Tecan GENios reader (Tecan, Männedorf, Switzerland) at 573 nm. Statistical significance was calculated with an unpaired t-test (two-tailed) using GraphPad Prism software (n = 2; * p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001).

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Cell Proliferation Assay Using MTT

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To assess cell proliferation, a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT)-based protocol was used. Directly after electroporation, cells were seeded at a density of 2500 (HUH6 and HepT1) or 5000 (HepG2) cells per well in a 96-well format (Nunc, Wiesbaden, Germany) in 100 μL RPMI medium. Ten microliters of MTT labeling agent (5 mg/mL, Sigma) was added to each well and incubated at 37 °C for 4 h. One hundred microliters of a SDS-HCl solution was added to each well and mixed thoroughly. Microplate was incubated overnight at 37 °C in a humidified chamber. The absorbance of this colorimetric reaction was quantified on the GENios reader (Tecan, Männedorf, Switzerland) by measuring at a wavelength of 595 nm.

Beck A., Trippel F., Wagner A., Joppien S., Felle M., Vokuhl C., Schwarzmayr T., Strom T.M., von Schweinitz D., Längst G, & Kappler R. (2018). Overexpression of UHRF1 promotes silencing of tumor suppressor genes and predicts outcome in hepatoblastoma. Clinical Epigenetics, 10, 27.

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Cathepsin B Activity Assay Protocol

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Cathepsin B activity assays were performed as described (Barrett, 1980 (link); Brix et al., 1996 (link); Mayer et al., 2009 (link)). In brief, protein samples prepared from thyroid tissues of n = 3 biological replicas per each genotype and age group, respectively, were assayed in triplicates by monitoring cleavage of 10 μM cathepsin B-specific substrate N-benzyloxycarbonyl-argininyl-arginine-7-amido-4-methylcoumarin hydrochloride (Z-Arg-Arg-AMC^∗HCl; Bachem, Bubendorf, Switzerland, #I-1135) at pH 6.0, and for 60 min at 40°C. In negative controls, prepared for each sample, 10 μM E-64 were added at the start of the reaction time. Substrate cleavage was stopped by the addition of 2 M Tris-HCl (pH 9.0). The amounts of released AMC were quantified by measuring the fluorescence with a Tecan GENios Reader (Tecan Deutschland GmbH) using an excitation wavelength of 360 nm and emission reading at 465 nm.

Qatato M., Szumska J., Skripnik V., Rijntjes E., Köhrle J, & Brix K. (2018). Canonical TSH Regulation of Cathepsin-Mediated Thyroglobulin Processing in the Thyroid Gland of Male Mice Requires Taar1 Expression. Frontiers in Pharmacology, 9, 221.

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Cell proliferation and cell-cycle assay

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The cells were seeded in 96-well culture plates at a density of 6 9 10 3 cells/well and treated with 10 nM siRNA. The CellTiter 96 Aqueous One Solution (Promega) assay was performed to determinate the gene downregulation effect on cell proliferation at 48, 72, and 96 h after transfection. For each well, 20 lL MTS was added and the plates were maintained 4 h at 37 °C. The absorbance was measured at 550 nm using a TECAN GENios reader. All experiments were effectuated in triplicate. Values were calculated as mean and SD of percentage change induced by each siR-NAs as compared to control cells. For the cell-cycle assay, the cells treated with siRNA for 72 h were fixed in 70% cold ethanol for at least 30 min at -20 °C. After washing twice in phosphate-buffered saline (PBS), cells were incubated 15 min at 37 °C with RNAse A (1 mg/ml) and 1 h with propidium iodide (100 lg/ml). The acquisition used a Epics Beckman Coulter flow cytometer. Data were analyzed using FlowJo software and expressed as fractions of cells in the different cell-cycle phases.

Chivu-Economescu M., Dragu D.L., Necula L.G., Matei L., Enciu A.M., Bleotu C, & Diaconu C.C. (2017). Knockdown of KRT17 by siRNA induces antitumoral effects on gastric cancer cells. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association, 20(6).

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Adenylate Kinase-Based Cell Viability Assay

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Adenylate kinase is an enzyme present in the media commonly used as an indicator of cell suffering and cell death [17 (link)]. Concentration of adenylate kinase in the medium from cultured neurons was measured using the ToxiLight BioAssay (Cambrex, Belgium, EU). 20 µm of medium were placed in a Luminunc™ plate well and 100 µl of the adenylate kinase detection kit were added to the well. After 5 min incubation at RT, luminescence was measured using a microplate luminescence Tecan Genios reader (Tecan, Switzerland) with an integrated read time of 1 s. A black divider was used to avoid cross-talking between wells and the value of a well without medium was used as reference.

Arnhold M., Dening Y., Chopin M., Arévalo E., Schwarz M., Reichmann H., Gille G., Funk R.H, & Pan-Montojo F. (2016). Changes in the sympathetic innervation of the gut in rotenone treated mice as possible early biomarker for Parkinson’s disease. Clinical Autonomic Research, 26, 211-222.

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Quantifying IL-1β in Microglia Cultures

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The levels of IL-1β in the culture media of the primary microglia and mixed glia were assessed using an enzyme-linked immunosorbent assay (ELISA) kit according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN, USA; Cat. DY501). Plates were read at a wavelength of 450 nm using a TECAN Genios reader (TECAN, Durham, NC, USA) and Magellan version 7.0 software.

Huang Y.C., Hsu S.M., Shie F.S., Shiao Y.J., Chao L.J., Chen H.W., Yao H.H., Chien M.A., Lin C.C, & Tsay H.J. (2022). Reduced mitochondria membrane potential and lysosomal acidification are associated with decreased oligomeric Aβ degradation induced by hyperglycemia: A study of mixed glia cultures. PLoS ONE, 17(1), e0260966.

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Cytokine Production by pDC upon Viral Infection

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40,000 highly-enriched pDC were incubated in wells of a 96-well round-bottom plate with a combination of T. gondii and HSV or HIV as described above, in a final volume of 200μL. Culture supernatants were collected 18 hrs post addition of virus and analyzed using the BenderMed Systems human IFN-a module set and the OptEIA® Human TNF-a Set. Absorbance was recorded at 450nm using TECAN GENios reader and concentration of cytokines was calculated using Microsoft Excel from a linear curve fit. For determination of IL-10 levels, samples were sent to Myriad RBM and analyzed for expression of inflammatory cytokines using Human InflammationMAP v.1.0.

Pierog P.L., Zhao Y., Singh S., Dai J., Yap G.S, & Fitzgerald-Bocarsly P. (2017). Toxoplasma gondii inactivates human plasmacytoid dendritic cells by functional mimicry of IL-10. Journal of immunology (Baltimore, Md. : 1950), 200(1), 186-195.

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Determining MIC of HU-210 on V. harveyi

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V. harveyi wild-type strain BB120 was grown at 30 °C in the presence of HU-210 at concentrations ranging from 2–4 mg/ml in 96-well transparent plates (Nunc, Roskilde, Denmark). After 24 h of incubation, bacterial growth was determined by absorbance at 595 nm (OD₅₉₅) (TECAN GENios reader, Schweiz, Austria). The MIC was recorded as the minimum concentration that exhibited visible growth inhibition and all further experiments were performed at sub-MIC [38 (link)].

Soni D., Smoum R., Breuer A., Mechoulam R, & Steinberg D. (2015). Effect of the synthetic cannabinoid HU-210 on quorum sensing and on the production of quorum sensing-mediated virulence factors by Vibrio harveyi. BMC Microbiology, 15, 159.

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