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Lb 940 luminometer

Manufactured by Berthold Technologies
Sourced in Germany, Switzerland

The LB 940 luminometer is a laboratory instrument used for the detection and measurement of luminescence. It is designed to quantify the light output from various types of luminescence-based assays and reactions. The LB 940 provides reliable and accurate luminescence data for a wide range of applications.

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7 protocols using lb 940 luminometer

1

Luciferase assay in RPTEC cells

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RPTEC cells were seeded into 24-well plates at a density of 1.5 x 105 cells/well. Cells were transfected using Lipofectamine 2000 (ThermoFisher scientific, Waltham, Massachusetts, USA), with 600 ng of DNA and 30 ng of a control vector for transfection efficiency, using the constitutive expression pCMV-Red Firefly Luc Vector (16156, ThermoFisher scientific, Waltham, MA, USA). After transfection, cells were exposed to 10 ng TNFα (< 0.1 endotoxin units per μg of protein, 210-TA, R&D Systems, Minneapolis, MN, USA) or the vehicle control for 24 h and then luciferase activity was determined using a Gaussia-firefly luciferase dual assay kit (16181, ThermoFisher scientific, Waltham, MA, USA) and a LB940 luminometer (Berthold, Germany).
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2

Placental Oxidative Stress Evaluation

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Placental tissue (200 mg) was homogenized. The reactive oxygen species (ROS) concentration analysis was performed by the chemiluminescence (CL) assay using luminol as an indicator of radical formation. The tissue samples were recorded at room temperature using an LB 940 luminometer (Berthold Technologies, Bad Wildbad, Germany) in the presence of enhancers as described previously [16 (link)]. The contents of malondialdehyde (MDA) were assayed using colorimetric methods with a spectrophotometer (Biomate 5, Thermo Electron Corporation, Rochester, NY, U.S.A.). Total antioxidant capacity (TAC) was measured with a commercial kit (Nanjing JianCheng Bioengineering Institute, A015, Nanjing, China) as per manufacturer’s instructions.
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3

Quantifying HCV Replication Levels

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We used four HCV subgenomic replicon cell lines, FLR3-1 (genotype 1b, Con-1)47 (link), R6FLR-N (genotype 1b, strain N)26 (link), JFH-1/FLR/K4 (genotype 2a)48 (link), and RMT-tri (genotype 1a)49 (link), which have the firefly luciferase gene for the sensitive and precise quantification of the HCV replication levels using a luciferase assay. We also used REF cells30 (link) which harbor the divided-full genome replicon for analysis of the HCV 5′ UTR sequences. Each cell line was seeded at a density of 5 × 103 per well in 96-well tissue culture plates, and grown (at 37°C and 5% CO2) in complete Dulbecco's modified Eagle's medium supplemented with Glutamax I (Invitrogen, Carlsbad, CA) and containing 5% fetal calf serum (Invitrogen). Cells were transfected with 30 nM siRNA using RNAiMax (Invitrogen, Carlsbad, CA). After 72 hours, luciferase activity was determined in triplicate using the Steady-Glo or Bright-Glo luciferase assay kit (Promega Madison, WI). The luciferase signal was measured using an LB940 luminometer (Berthold, Freiburg, Germany) and the results were expressed as the mean percentage of control. IC50 values of siRNA were calculated by nonlinear curve-fitting using the equation: Y = 100-(YBottom × X/(IC50 + X)), where Y represents percent inhibition and X represents the concentration of siRNAs.
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4

Chemiluminescence Assay on PLB-985 Cells

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Chemiluminescence assay on differentiated PLB-985 WT, PLB-985 X-CGD, and PLB-985 NCF1 ΔGT cells, as well as transduced PLB-985 NCF1 ΔGT was conducted in 96-well plate format at a cell density of 1 * 105 cells/200 ml in a Mithras LB 940 Luminometer (Berthold Technologies GmbH, Zug, Switzerland) as described recently11 (link).
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5

Dual-Luciferase Reporter Assay for Transcriptional Activity

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Cells were seeded on a 24-well plate, and transfected with siRNAs. After 48 h, cells were transfected (GeneCopoeia) with 500 ng TOP flash or NF-κB reporter and 10 ng pRL-TK (Promega, Madison, WI, USA) plasmids using EndoFectin™-Plus. Assays were performed in accordance with the dual-luciferase assay specifications (Promega) using the Mithras LB 940 luminometer (Berthold, Bad Wildbad, Germany). The activity of firefly luciferase was normalized to measure the transfection efficiency. All experiments were performed at least 3 times.
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6

Mitochondrial dehydrogenase activity assay

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After treatments, succinate dehydrogenase (SD) activity assay (MTT) [11 (link)] was applied to HEK293 cells as described previously [12 (link)]. The integrity of mitochondrial dehydrogenase enzymes indirectly reflects the cellular mitochondrial respiration. The optical density was measured at 570 nm using a Mithras LB 940 luminometer (Berthold, Thoiry, France).
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7

Luciferase Assay for Flg22-Induced Response

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Luciferase reporter gene assay was conducted following the method of Zheng et al. (2014). Protoplasts were treated with/without flg22 to a final concentration of 500 nM. The luminescence reflecting the luciferase activity was measured at different time‐points using a Mithras LB 940 luminometer (Berthold, Bad Wildbad, Germany). Luciferase activity (+/−flg22) was normalized by measuring β‐glucuronidase (GUS) activity upon lysis with an equal volume of CCLR solution and incubation of 10 μl protoplast extract with 90 μl MUG substrate.
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