Hepg2 lucia ahr reporter cells

Manufactured by InvivoGen

Sourced in United States, France

The HepG2-Lucia™ AhR reporter cells are a genetically modified human hepatocellular carcinoma cell line that stably expresses the Lucia luciferase reporter gene under the control of an aryl hydrocarbon receptor (AhR) responsive promoter. The cells can be used to measure the activation of the AhR signaling pathway by various compounds.

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Lab products found in correlation

Spectramax m5, by Molecular Devices (2 mentions) Quanti luc, by InvivoGen (2 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions) Quanti luc reagent, by InvivoGen (2 mentions) Cytotox 96 non radioactive cytotoxicity assay, by Promega (2 mentions) 96 well flat bottom plate, by Corning (1 mentions) Penicillin streptomycin p s p4333, by Merck Group (1 mentions) Dmem glutamax, by Thermo Fisher Scientific (1 mentions) Caco 2 cell line, by American Type Culture Collection (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Ht 29 cell line, by American Type Culture Collection (1 mentions) Ht29 lucia, by InvivoGen (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) α mem, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Normocin, by InvivoGen (1 mentions) Zeocin, by InvivoGen (1 mentions)

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HepG2-Lucia™ Ahr Cells (2 citations)

5 protocols using hepg2 lucia ahr reporter cells

Measuring AhR Activation using Luciferase Assay

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The AhR activity was measured using a luciferase reporter assay method as described previously.^[³⁵ (link)
^] In brief, HepG2‐Lucia AhR reporter cells (InvivoGen, San Diego, CA) were grown and maintained according to the manufactures instructions. To induce AhR, 20 µL samples separately from three independent replicates and 180 µL cell suspension containing about 20 000 cells were added per well in triplicate to a flat‐bottom 96‐well plate (Corning, USA), which diluted samples by ten times. The plate was placed at 37 °C in a CO₂ incubator for 48 h, and then 20 µL of stimulated cells supernatant was transferred into a white walled clear bottom 96‐wells plate (Corning, USA), followed by addition of 50 µL QUANTI‐Luc (InvivoGen). The luminescence was immediately measured by a Spectramax M5 (Molecular Devices, USA). The results are expressed as the percentage of AhR activity of positive control (5 µm β‐naphthoflavone or β‐NAPH in DMSO) and normalized by either DMSO, cell growth medium or control samples fermented without substrates.

Huang Z., Schoones T., Wells J.M., Fogliano V, & Capuano E. (2021). Substrate‐Driven Differences in Tryptophan Catabolism by Gut Microbiota and Aryl Hydrocarbon Receptor Activation. Molecular Nutrition & Food Research, 65(13), 2100092.

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Cell Culture Protocols for Cancer and Immune Cells

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The human colon adenocarcinoma HT-29 cell line was obtained from the American Type Culture Collection (ATCC). Cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with glutaMAX™ (Gibco), 10% (v/v) fetal bovine serum (FBS; Gibco) and 0.01% (v/v) penicillin/streptomycin (P/S, P4333; Sigma). The cultures were grown at 37 °C and 5% CO₂ until 80% confluence was reached.
The Caco-2 cell line was obtained from the ATCC^® and maintained in DMEM glutaMAX™ (Gibco), 20% heat-inactivated FBS, 1% non-essential amino acids (Gibco) and 0.01% (v/v) P/S. Cells were grown at 37 °C under conditions of 10% CO₂ until 80% confluence was reached.
HepG2-Lucia™ AhR reporter cells were obtained from Invivogen (San Diego, CA, USA). Cells were maintained according to the manufacturer’s protocol.
Human peripheral blood mononuclear cells (PBMCs) isolated from the blood of healthy donors were obtained from Stemcells (Vancouver, BC, Canada) and stored in liquid nitrogen until use.

Maillard F., Meynier M., Mondot S., Pepke F., Galbert C., Torres Maravilla E., Kropp C., Sokol H., Carvalho F.A., Jacouton E., Holowacz S., Langella P., Chain F, & Martín R. (2023). From In Vitro to In Vivo: A Rational Flowchart for the Selection and Characterization of Candidate Probiotic Strains in Intestinal Disorders. Microorganisms, 11(4), 906.

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Measurement of AhR Activity in Animal Stool

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The AhR activity of animal stool samples was measured using HepG2-Lucia™ AhR reporter cells (InvivoGen, France). Cells were seeded into a 96-well plate and stimulated with animal stool samples or different amounts of tryptophol for 24 hours. Luciferase activity was measured using a luminometer and Quanti-Luc reagent (InvivoGen). The results were normalized on the basis of the negative luciferase activity of the control, cytotoxicity measurement (CytoTox 96 Non-radioactive Cytotoxicity Assay, Promega) and feces weight.

Meynier M., Baudu E., Rolhion N., Defaye M., Straube M., Daugey V., Modoux M., Wawrzyniak I., Delbac F., Villéger R., Méleine M., Borras Nogues E., Godfraind C., Barnich N., Ardid D., Poirier P., Sokol H., Chatel J.M., Langella P., Livrelli V., Bonnet M, & Carvalho F.A. (2022). AhR/IL-22 pathway as new target for the treatment of post-infectious irritable bowel syndrome symptoms. Gut Microbes, 14(1), 2022997.

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AhR Activity Measurement in Cell Lines

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The AhR activity was measured using HepG2-Lucia™ AhR reporter cells (InvivoGen, France) and HT29-Lucia™ AhR reporter cells. Cells were seeded into a 96-well plate and stimulated with for 24 hours according to the manufacturer’s protocol. Luciferase activity was measured using a luminometer and Quanti-Luc reagent (InvivoGen). The results were normalized on the basis of the negative luciferase activity of the control and cytotoxicity measurement (CytoTox 96 Non-radioactive Cytotoxicity Assay, Promega).

Modoux M., Rolhion N., Lefevre J.H., Oeuvray C., Nádvorník P., Illes P., Emond P., Parc Y., Mani S., Dvorak Z, & Sokol H. (2022). Butyrate acts through HDAC inhibition to enhance aryl hydrocarbon receptor activation by gut microbiota-derived ligands. Gut Microbes, 14(1), 2105637.

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Measuring Aryl Hydrocarbon Receptor Activity

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HepG2-Lucia™ AhR reporter cells (InvivoGen) were grown according to the manufactures instructions in Minimum Essential Medium Eagle -Alpha Modification (α-MEM; Gibco USA), 10% (v/v) heat inactivated fetal calf serum (Gibco), 1× non-essential amino acids (Gibco), Penicillin-Streptomycin (100 U mL -1 -100 µg mL -1 ; Gibco), 100 µg mL -1 Normocin™ (InvivoGen) and 100 µg mL -1 Zeocin™ (InvivoGen). To measure AhR activation, 20 μl sample was added in triplicate to a 96-wells plate (Corning, USA). Thereafter, 180 μl cell suspension (∼20 000 cells per well) was added to each well and the plate was incubated for 48 hours at 37 °C in a CO 2 incubator. After incubation, 20 μL stimulated cell supernatant was transferred into a white walled clear bottom 96-wells plate (Corning), followed by addition of 50 μL QUANTI-Luc™ (InvivoGen). Luminescence was measured immediately using a Spectramax M5 (Molecular devices, USA). AhR activation was expressed as percentage of the activity of the positive control (5 μM β-naphthoflavone in DMSO). Results of AhR activation were corrected for their corresponding negative controls, either DMSO or medium.

Koper JEB ., Kortekaas M ., Loonen LMP ., Huang Z ., Wells JM ., Gill CIR ., Pourshahidi LK ., McDougall G ., Rowland I ., Pereira-Caro G ., Fogliano V , & Capuano E . (2020). Aryl hydrocarbon Receptor activation during in vitro and in vivo digestion of raw and cooked broccoli (brassica oleracea var. Italica). Food & function, 11(5).

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