Cell viability was monitored as previously described [34 (link)]. Briefly, 100 μL of cultures diluted to a final OD600 = 0.025 ± 0.0025 were placed into 96-well microplates; 10 μL of cell titer blue reagent (Promega, WI, USA) were added, and the plates were incubated for 3 h at 30 °C. Fluorescence was measured in 30 min intervals at emission wavelength 580 nm using the Biotek Power Wave XS Microplate Spectrophotometer (Biotek® Instruments, Winooski, VT, USA). Doses of chicory SFE extracts ranging from 100 to 1000 µg/mL, and purified fractions containing 8-deoxylactucin and 11β,13-dihydro-8-deoxylactucin in the range of 12.5–100 µg/mL were tested in the viability assays. Readings were performed using a Biotek Power Wave XS plate spectrophotometer (Biotek® Instruments, Winooski, VT, USA).
Biotek power wave xs microplate spectrophotometer
Manufactured by Agilent Technologies
Sourced in United States
The BioTek Power Wave XS Microplate Spectrophotometer is a versatile instrument designed for absorbance-based assays in microplates. It measures the absorption of light at various wavelengths to quantify analytes and assess the concentration of substances within samples.
Automatically generated - may contain errors
Lab products found in correlation
Celltiter blue reagent, by Promega (1 mentions)
Edta coated vacutainer tubes, by BD (1 mentions)
Aprotinin, by Merck Group (1 mentions)
Ezrmgh 45k, by Merck Group (1 mentions)
Ezrgra 90k, by Merck Group (1 mentions)
Vitros analyzer, by Ortho Clinical Diagnostics (1 mentions)
P hydroxymercuribenzoic acid, by Merck Group (1 mentions)
2 protocols using biotek power wave xs microplate spectrophotometer
1
Cell Viability Assay with Chicory Extracts
2
Plasma Biomarker Extraction and Quantification
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Blood was collected into 3 mL ice-cold EDTA-coated vacutainer tubes (BD Biosciences, Franklin Lakes, NJ) and aliquoted into microfuge tubes containing either the protease inhibitors p-hydroxymercuribenzoic acid (Sigma Aldrich; final concentration 1 mM; used for ghrelin measurement) or aprotinin (Sigma Aldrich; final concentration 250 KIU/mL; used for LEAP2 measurement) or lacking protease inhibitor (used for GH, IGF-1, liver enzymes, total cholesterol, and triglycerides). The samples were immediately centrifuged at 4 °C at 1,500 g for 15 min. To stabilize the acyl-group on ghrelin, 1N hydrochloric acid was added to the p-hydroxymercuribenzoic acid-treated plasma to achieve a final concentration of 0.1 N. Processed samples were stored at −80 °C in small aliquots until further analysis of plasma hormone levels. ELISA kits were used for acyl-ghrelin (#EZRGRA-90K; Millipore-Merck, MA), GH (#EZRMGH-45K; Millipore-Merck), IGF-1 (#80574; Crystal Chem, IL), and LEAP2 [#EK-075-40; Phoenix Pharmaceuticals, Inc., CA]. Colorimetric assays were performed using a BioTek PowerWave XS Microplate spectrophotometer (BioTek, Winooski, VT) using BioTek KC4 junior software. Measurements of plasma AST, ALT, total cholesterol and triglycerides were measured with a VITROS analyzer (Ortho Clinical Diagnostics) in the UT Southwestern Mouse Metabolic Phenotyping Core.
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