Fluostar omega microplate spectrophotometer

Manufactured by BMG Labtech

Sourced in United States

The FLUOstar Omega is a microplate spectrophotometer manufactured by BMG LABTECH. It is designed to measure the absorbance and fluorescence of samples in microplates. The instrument can perform various photometric measurements and is suitable for a wide range of applications in life science research and diagnostics.

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Lab products found in correlation

Sheep erythrocytes, by Rockland Immunochemicals (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Celltiter blue reagent, by Promega (1 mentions) Lumistar omega microplate luminometer, by BMG Labtech (1 mentions) Tnf α, by Thermo Fisher Scientific (1 mentions) 6 well plate, by BD (1 mentions) Tnf α, by R&D Systems (1 mentions) Las 4000, by Fujifilm (1 mentions) Sheep blood, by Hardy Diagnostics (1 mentions) Millex gp, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Luria bertani lb, by Merck Group (1 mentions)

5 protocols using fluostar omega microplate spectrophotometer

Inflammasome-Activated Caspase-1 Assay

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For each sample, an equal volume of supernatant was transferred into 96-well plates and equilibrated at room temperature for 5 min. To detect inflammasome-related caspase-1 activity, an equal volume of the Caspase-Glo Inflammasome Reagent (Promega, Madison, WI, USA) was added in each well according to the manufacturer’s instructions. The plate was incubated at room temperature for 1 h in the dark and the relative luminescence units (RLU) of assays were determined in the LUMIstar Omega microplate luminometer (BMG Labtech, Ortenberg, Germany) according to the manufacturer’s directions.
For cell viability, cells were incubated with Cell Titer Blue Reagent (Promega, Madison, WI, USA) at 37 °C for 1 h according to the manufacturer’s instruction. The relative fluorescence (560Excitation/590Emission) was detected with the FLUOstar Omega microplate spectrophotometer (BMG Labtech, Ortenberg, Germany) following the manufacturer’s instructions.

Scalavino V., Piccinno E., Valentini A.M., Schena N., Armentano R., Giannelli G, & Serino G. (2023). miR-369-3p Modulates Intestinal Inflammatory Response via BRCC3/NLRP3 Inflammasome Axis. Cells, 12(17), 2184.

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Cytokine and Phospho-RTK Profiling of Beta-TC-6 Cells

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Beta-TC-6 cells with and without Dsg2 knockdown were seeded into 6 well plates (Falcon). After 48 h, the media was aspirated, and cells were either treated without or with TNFα (100 ng/ml, Life Technologies) for a further 24 h. Conditioned media was then collected and centrifuged at 1500 rpm for 5 min, the supernatant was separated and stored at −80 °C until use. Cells were lysed directly in the well with lysis buffer (provided and pre-prepared with protease inhibitors). Lysates were collected and clarified by centrifugation (13,000 rpm, 8 min, 4 °C). Protein concentration was determined using a Pierce™ BCA protein assay kit (Thermo Fisher Scientific) according to the manufacturer’s instructions, and the protein concentration was measured at 540 nm using the FLUOstar Omega Microplate Spectrophotometer (BMG Labtech, Mornington, VIC, Australia). The cytokine and phospho-receptor tyrosine kinase (RTK) profile of Beta-TC-6 cells (± siDSG2 and ± TNFα treatment) was assessed using the Proteome Profiler Mouse Array Kits (R&D Systems) according to manufacturer’s instructions. The membrane was visualised using the LAS-4000 (FujiFilm, Tokyo, Japan). Pixel density quantification of the dots on the membrane was performed using ImageJ Fiji software.

Myo Min K.K., Rojas-Canales D., Penko D., DeNichilo M., Cockshell M.P., Ffrench C.B., Thompson E.J., Asplund O., Drogemuller C.J., Prasad R.B., Groop L., Grey S.T., Thomas H.E., Loudovaris T., Kay T.W., Mahoney M.G., Jessup C.F., Coates P.T, & Bonder C.S. (2022). Desmoglein-2 is important for islet function and β-cell survival. Cell Death & Disease, 13(10), 911.

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Hemolytic Activity Quantification of B. thuringiensis

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WT and ΔslpA B. thuringiensis were cultured for 18 hours as mentioned above, then centrifuged at 3000g for 15 minutes. Supernatants were then filter sterilized (0.22 μm; Millex-GP, Merck Millipore Ltd., Cork, Ireland) and serially diluted 1:2 in PBS (pH 7.4). Diluted supernatants were incubated 1:1 with 4% (vol/vol) sheep erythrocytes (Rockland Immunochemicals, Pottstown, PA, USA) in a 96-well round bottom plate for 30 minutes at 37°C. The plate was centrifuged at 1892g for 10 minutes to remove the unlysed erythrocytes.
The supernatants were carefully transferred into a 96-well, flat-bottom plate and hemoglobin release was measured spectrophotometrically at 490 nm by using a FLUOstar Omega microplate spectrophotometer (BMG Labtech, Cary, NC, USA). Values are expressed as the percent hemolysis relative to a 100% lysis control in which 5% rabbit erythrocytes were lysed in double-distilled H₂O.23 (link),27 (link),48 (link),76 ,77 (link) Values represent the mean results ± SEM of two independent experiments. These strains were also incubated on tryptic soy agar (TSA) supplemented with 5% sheep blood (Hardy Diagnostics, Santa Maria, CA, USA) for 18 hours at 37°C, and colony morphology and hemolytic phenotypes were compared.

Mursalin M.H., Coburn P.S., Livingston E., Miller F.C., Astley R., Fouet A, & Callegan M.C. (2019). S-layer Impacts the Virulence of Bacillus in Endophthalmitis. Investigative Ophthalmology & Visual Science, 60(12), 3727-3739.

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Hemolysis Assay for Phage Lysin and Antibiotic

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Phage lysin PlyB (125 µg/mL), filter-sterilized B. cereus Sup, and the antibiotic GAT (250 µg/mL) were serially diluted 1:2 in PBS (pH 7.4) in a 96-well round bottom plate. Diluted PlyB, GAT, and B. cereus Sup were incubated 1:1 with 4% (vol/vol) sheep erythrocytes (Rockland Immunochemicals, Pottstown, PA, USA) for 30 minutes at 37°C. Unlysed erythrocytes were removed by centrifuging the plate at 1,892 g for 10 minutes. The supernatants were carefully transferred into a 96-well flat bottom plate, and hemoglobin release was measured spectrophotometrically at 490 nm using a FLUOstar Omega microplate spectrophotometer (BMG Labtech, Cary, NC, USA). Values are expressed as the percent hemolysis relative to a 100% lysis control in which 5% rabbit erythrocytes were lysed in double-distilled water (56 (link), 57 (link)).

Mursalin M.H., Astley R., Coburn P.S., Bagaruka E., Hunt J.J., Fischetti V.A, & Callegan M.C. (2023). Therapeutic potential of Bacillus phage lysin PlyB in ocular infections. mSphere, 8(4), e00044-23.

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Bacillus cereus Phage Lysin PlyB Evaluation

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B. cereus ATCC 14579 was grown overnight in brain–heart infusion medium (BHI; VWR, Radnor, PA, USA). Overnight B. cereus cultures were centrifuged, washed, and resuspended in phosphate buffered saline (PBS, pH 7.4) to an OD₆₀₀ of 2.00. Diluted bacteria (50 µL) were plated in a U-bottomed 96-well plate in triplicate. B. cereus phage lysin PlyB (250 µg/mL) was added and serially diluted to a range of concentrations from 125 to 0 μg/mL. PlyB dilution plates were then incubated at 37°C in a FLUOstar Omega microplate spectrophotometer (BMG Labtech, Cary, NC, USA) for 60 minutes. OD₆₀₀ was measured at 0.5, 2.5, 5, 10, 20, 30, 45, and 60 minutes (53 (link)).
PlyB activity in different growth conditions was also assessed by turbidity assay. An overnight B. cereus culture was centrifuged, washed, and resuspended in BHI, Luria-Bertani (LB; Sigma, St Louis, MO, USA), Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Grand Island, NY, USA), Vit (PelFreez, Rogers, AR, USA), or human plasma-like media (Gibco) to an OD₆₀₀ of 2.00. An amount of 50 µL of the PBS, BHI, LB, DMEM, Vit, and plasma suspensions was plated in triplicate in a U-bottomed 96-well plate. An amount of 50 µL B. cereus phage lysin PlyB (250 µg/mL) was added to each well to a final concentration of 125 µg/mL and an OD₆₀₀ of 1.00. Plates were then incubated, and OD₆₀₀ was measured as above.

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