Cell viability and proliferation was determined more than 3 times using 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) reagent (Invitrogen, Carlsbad, CA, USA). Briefly, cells were placed in 96-well plates and incubated for 24 hr at 37℃ under 5% CO2, at which point 100 µL of 0.5 mg/mL MTT solution was added to each well and incubated for 4 hr at 37℃. Formazan absorbance was read at 490 nm using a GENios ELISA reader (TECAN, Mannedorf, Switzerland).
Genios elisa reader
Manufactured by Tecan
Sourced in Austria, Switzerland
The GENios ELISA reader is a laboratory instrument designed for performing enzyme-linked immunosorbent assay (ELISA) experiments. It is capable of measuring the absorbance of samples in microplate format, which is a key step in ELISA procedures.
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Lab products found in correlation
7 protocols using genios elisa reader
1
MTT Assay for Cell Viability
2
XTT Cell Viability Assay Protocol
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The viability of cultured cells was examined using the XTT assay (Roche, Basel, Switzerland) according to the manufacturer’s instructions. In brief, treated RINm5F cells were cultured in 96-well plates (20,000 cells per well) in 100 μL of medium. After 24 h or 48 h, 50 μL of the XTT labeling mixture was added to each well to a final XTT labeling reagent concentration of 0.3 mg/mL with an electron coupling reagent. After incubation at 37 °C for 8 h, the chemiluminescence intensity was assessed at 450 nm using a GENios ELISA reader (Tecan, Chapel Hill, NC, USA).
3
Serum CDT Quantification by ELISA
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A quantitative, sandwich enzyme immunoassay technique was used to measures CDT from serum samples.[19 ] The assay was performed by ELISA (Enzyme-Linked Immuno-Sorbent Assay) technique (Tecan GENios ELISA reader, Austria GmbH, Austria), using Magellan software. The procedure was followed as per the protocol provided by the manufacturer (Cusabio, USA). The concentration of CDT was determined using the professional software “Curve Expert” to make a standard curve from the web (www.cusabio.com ). The levels obtained in direct serum samples were compared with their corresponding serum samples spotted on to filter paper.
4
Screening and Confirmation of Analytes
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Primary screening was performed by enzyme-linked immunosorbent assay (ELISA) technique (Tecan GENios ELISA reader, Austria GmbH, Austria) using Magellan software. Further confirmation was done on gas chromatograph model 7890A, Agilent India Pvt. Ltd., USA, equipped with 7683B series Autosampler; split/splitless inlet, fused silica capillary column coated with HP-5 cross-linked 5% diphenyl and 95% dimethylpolysiloxane (30 m × 0.320 i.d., 0.25 μm film thickness); and nitrogen phosphorus detector (NPD) with electronic pneumatic control. System control, data acquisition, and analysis were performed with gas chromatography (GC) ChemStation G2075BA software.
5
Hematological and Hormonal Biomarker Analysis
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Fasting venous whole blood samples (5 ml) were collected in tubes with EDTA to analyze hematological profiles (i.e. neutrophils and lymphocytes) using an automated hematology analyzer (Sysmex XT-2000, Sysmex Corp., Kobe, Japan) in the light of manufacturer’s instruction. The remaining whole blood samples were centrifuged at 4°C, 3000 rpm for 10 min to obtain plasma samples. The plasma samples were then stored at -80°C until analyzed. Cortisol and dehydroepiandrosterone-sulfate (DHEA-S) were evaluated using commercially available enzyme-linked immunosorbent assay (ELISA) kits (Cayman Chemical Co., Ann Arbor, MI, USA and IBL Inc., Hamburg, Germany) according to the manufacturer’s instructions. All ELISA assays were read using a TECAN Genios ELISA reader (Salzburg, Austria). The optical density was used to determine the amount of each hormone in these plasma samples.
6
Plasma Thyroid Hormone Quantification
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Plasma L-thyroxine (T4) and 3,5,3′ tri-iodothyronine (T3) quantitative measurements were performed with EIAT3C T3andEIAT4CT4ELISA kits (ThermoScientific LSG, MA, USA), according to manufacturer’s instructions. L-thyroxine and 3,5,3′ triiodothyronine levels were expressed as ng/mL of plasma. Measurements were performed with Genios ELISA reader (Tecan, Männedorf, Switzerland).
7
Enzyme and Gas Chromatographic Analysis
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Screening was performed by enzyme-linked immunosorbent assay (ELISA) technique (Tecan GENios ELISA reader, Austria GmbH, Austria) using Megellan Software V1.30 (Tecan Austria, Austria).
Further confirmation of the samples was done using gas chromatograph (model 7890A, Agilent India Pvt. Ltd, USA). The gas chromatography (GC) was equipped with 7683B series Auto Sampler (split/split-less inlet, fused silica capillary column coated with HP-5 cross-linked 5% diphenyl, and 95% dimethylpolysiloxane (30 m × 0.320 i.d., 0.5 μm film thickness). The temperature gradient used was 1 min at 170°C, 10°C/min from 170°C to 270°C, and 5 min at 270°C. Nitrogen was used as a carrier gas at a flow rate of 10 ml/min. The injector temperature was 280°C. The injection volume was 2 μl for each GC run, and the split ratio was kept 1:10. The nitrogen phosphorus detector (NPD) was used with electronic pneumatic control at 300°C. System control, data acquisition, and analysis were performed with GC Chem Station G2075BA software.
Further confirmation of the samples was done using gas chromatograph (model 7890A, Agilent India Pvt. Ltd, USA). The gas chromatography (GC) was equipped with 7683B series Auto Sampler (split/split-less inlet, fused silica capillary column coated with HP-5 cross-linked 5% diphenyl, and 95% dimethylpolysiloxane (30 m × 0.320 i.d., 0.5 μm film thickness). The temperature gradient used was 1 min at 170°C, 10°C/min from 170°C to 270°C, and 5 min at 270°C. Nitrogen was used as a carrier gas at a flow rate of 10 ml/min. The injector temperature was 280°C. The injection volume was 2 μl for each GC run, and the split ratio was kept 1:10. The nitrogen phosphorus detector (NPD) was used with electronic pneumatic control at 300°C. System control, data acquisition, and analysis were performed with GC Chem Station G2075BA software.
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