The immobilization of the bioreporter bioluminescent bacteria cells was examined on several different surfaces. The filter membrane disks in a diameter of 0.5 mm were created by punching into parafilm and polyester membranes. Then, a 50 μL bacterial suspension with or without additives (100 μg/mL ampicillin and 10 mM glucose) at different concentrations was immobilized onto the filter membrane surface disks by incubation in petri dishes in an incubator at 37 °C for 2 h, until the solution fully evaporated. The filter membrane disks with the immobilized bacteria were subsequently ready for the monitoring of toxicants in liquid or long-term storage at room temperature. The bacterial bioluminescence activity was measured in a SynergyHTX multi-mode reader (BioTek Instruments Inc., Winooski, VT, USA), by placing it in white opaque 96-well microtiter plates (Nunc, Roskilde, Denmark). The plates containing the bacteria immobilized surface disks were exposed to different concentrations of toxicants in LB and control samples (n ≥ 3 for each tested condition). During the measurement, the temperature of the samples was maintained at 26 °C. The luminescence values are presented in relative light units (RLU).
White opaque 96 well microtiter plates
Manufactured by Thermo Fisher Scientific
Sourced in United States
White opaque 96-well microtiter plates are laboratory equipment designed for various assays and experiments. These plates have 96 individual wells arranged in a 8x12 grid format. The plates are made of a white opaque material, which helps to minimize well-to-well optical interference and improves signal-to-noise ratio in applications such as absorbance and fluorescence measurements.
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Lab products found in correlation
3 protocols using white opaque 96 well microtiter plates
1
Bioluminescent Bacteria Immobilization for Toxicant Monitoring
2
Bioluminescent Substrate Profiling of Human Liver Microsomes
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Triton-X100 was from Leagene (Beijing, China); Tris-HCl, from AKZ-Biotech (Tianjin, China); potassium chloride, ammonium bicarbonate, potassium dihydrogen phosphate and potassium hydrogen phosphate were from Jiangtian Chemical (Tianjin, China); glycerol was from Dingguo (Tianjin, China); 1 X PBS buffer was from Corning (Manassas, VA); human liver microsomes (HLMs) were from Sekisui XenoTech (Kansas City, KS, USA); Luciferin 6′ 2-fluorobenzyl ether (Luciferin-2FBE), Luciferin 6′ 3-fluorobenzyl ether (Luciferin-3FBE), Luciferin 6′ 2-furfuryl ether (Luciferin-2FE), Luciferin 6′ 3-furfuryl ether (Luciferin-3FE), Luciferin 6′ 2-(5-trifluoromethyl)furfuryl ether (Luciferin-TFM2FE), and Luciferin 6′ 3-thenyl ether (Luciferin-3TE) were synthesized as described [13 (link)]; the NADPH regeneration system (20X solution A: 26 mM NADP+, 66 mM glucose-6-phosphate, 66 mM MgCl2; and 100X solution B: 40 U/ml glucose-6-phosphate dehydrogenase in 5 mM sodium citrate, pH 5.5), Luciferin detection reagent (Cat# V8921), and Luciferin detection reagent with esterase (Cat# V8931) were from Promega (Madison, USA); white opaque 96-well microtiter plates were from Nunc (Thermofisher scientific, Lagenselbold, Germany); all other chemicals and reagents used were of the highest grade available.
3
Luminescence Assay for Proluciferin Compounds
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Proluciferin compounds (except luciferin-3FEME) and the NADPH regeneration system (20X solution A: 26 mM NADP+, 66 mM glucose-6-phosphate, 66 mM MgCl2; and 100X solution B: 40 U/ml glucose-6-phosphate dehydrogenase in 5 mM sodium citrate, pH 5.5), the luciferin detection reagent (Cat# V8921), and the luciferin detection reagent with esterase (Cat# V8931) were from Promega (Madison, WI). Luciferin 6' 3-furfuryl ether methyl ester (luciferin-3FEME) was synthesized as described [11 (link)]. Triton X-100 was from Leagene (Beijing, China), and Tris-HCl was from AKZ-Biotech (Tianjin, China). Potassium chloride, ammonium bicarbonate, potassium dihydrogen phosphate, and potassium hydrogen phosphate were from Jiangtian Chemical (Tianjin, China), and 1X TBS buffer, 1X PBS (0.1 mM CaCl2 (anhydrous), 2.7 mM KCl, 1.5 mM KH2PO4, 0.5 mM MgCl2, 137 mM NaCl, 8.1 mM Na2HPO4, pH 7.4) buffer were from Corning (Manassas, VA). White opaque 96-well microtiter plates were from Nunc (ThermoFisher Scientific, Lagenselbold, Germany). All other chemicals and reagents used were of the highest grade available.
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