N luminometer

Manufactured by Promega

Sourced in United States

The 20/20n Luminometer is a compact and versatile instrument designed for luminescence-based assays. It provides accurate and sensitive measurement of luminescent signals, making it suitable for a variety of applications in research and laboratory settings. The 20/20n Luminometer is capable of detecting and quantifying luminescent signals, enabling researchers to obtain reliable data for their luminescence-based experiments.

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Lab products found in correlation

Luciferase assay system, by Promega (7 mentions) Dual luciferase reporter assay system, by Promega (6 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (6 mentions) Passive lysis buffer (plb), by Promega (4 mentions) Reporter lysis buffer, by Promega (3 mentions) Ingenio electroporation kit, by Mirus Bio (2 mentions) Bradford method, by Bio-Rad (2 mentions) Adenosine 5 triphosphate bioluminescent somatic cell assay kit, by Merck Group (2 mentions) Luciferase assay kit, by Promega (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions) Amaxa basic nucleofector kit for primary mammalian fibroblasts, by Lonza (1 mentions) Amaxa cell line nucleofector kit 5, by Lonza (1 mentions) Nucleofector 2, by Lonza (1 mentions) Dual luciferase reporter assay kit, by Promega (1 mentions) Dual glotm luciferase assay system, by Promega (1 mentions) Ad nfkb luc, by Vector Biolabs (1 mentions) β galactosidase enzyme assay, by Promega (1 mentions)

Close spelling variants of this product

20/20n Single Tube luminometer

38 protocols using n luminometer

Glucose and Lactate Assays in U87MG Cells

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Glucose Uptake-Glo^TM and Lactate-Glo ^TM (Promega, Madison, WI, USA) assay kits were used to perform the glucose uptake and lactate secretion assays, respectively, as per the manufacturer’s instructions. Both assays were performed with or without FBS with similar results. For glucose uptake assays, log-phase U87MG cells were treated with 1 μM NVP-BEZ235 for 48 h under starvation conditions (without FBS). After treatment, 36,000 cells were washed with PBS and incubated with 2-deoxy-D-glucose (2DG) for 10 min at room temperature. Luminescence was recorded using a 20/20n luminometer from Turner Biosystems (Sunnyvale, CA, USA). Samples were examined in quadruplicate in replicate experiments.
For lactate secretion assays, log-phase U87MG cells were treated with 1 μM NVP-BEZ235 for 48 h under starvation conditions (without FBS). Then, 3 μL of culture media were placed into 97 μL of PBS and were incubated with the Lactate Detection Reagent for 60 min at room temperature. The luminescence was recorded using the 20/20n luminometer from Turner Biosystems. Cell numbers were counted for each sample and were used to normalize luminescence readings for comparisons. Samples were examined in quadruplicate in replicate experiments.

Udawant S., Litif C., Lopez A., Gunn B., Schuenzel E, & Keniry M. (2021). PI3K Pathway Inhibition with NVP-BEZ235 Hinders Glycolytic Metabolism in Glioblastoma Multiforme Cells. Cells, 10(11), 3065.

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Quantifying Cell Surface Kv1.3 Expression

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Analysis of surface expression of K_v1.3 was performed as previously described [43 (link),44 (link)]. Briefly, cells were fixed in 4% paraformaldehyde (20 min), washed, and blocked with fetal bovine serum (1%). Cells were then probed with anti-K_v1.3 primary antibody specific to an extracellular epitope (1:50, Alomone) and a secondary antibody conjugated to horse radish peroxidase. Chemiluminescence was detected using a 20/20ⁿ Luminometer (Turner Biosystems, Sunnyvale, CA) following application of SuperSignal West Femto Maximum Sensitivity substrate (Thermo Scientific) to cells. Plates were scraped and whole cell lysates were collected in RIPA lysis buffer, and quantified using a BCA assay (Thermo Scientific). Luminometry results were then normalized to protein content (μg/μL) and then to control to permit simple comparisons. Experiments were repeated at least three times. Control experiments using an antibody specific to actin (Sigma) in conjunction with a secondary antibody conjugated to horse radish peroxidase (the same as that used for K_v1.3 experiments) did not result in greater luminescence than using the secondary antibody alone, indicating that we were measuring surface-specific expression of K_v1.3 protein.

Silver K., Littlejohn A., Thomas L., Marsh E, & Lillich J.D. (2015). Inhibition of Kv channel expression by NSAIDs depolarizes membrane potential and inhibits cell migration by disrupting calpain signaling. Biochemical pharmacology, 98(4), 614-628.

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Quantifying Extracellular ATP Release

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ATP release from the cultured cell lines was measured using a commercially available rLuciferase/Luciferin (rL/L) reagent assay (Promega Enliten). Briefly, the samples were neutralized to pH 7.4 with 10 μL 4M Tris and were aliquoted to a new tube with 90 μL ATP-free water. Luciferase reagent was added 1 s before measurement in the 20/20n Luminometer Turner BioSystems (Sunnyvale, CA, USA). An ATP standard curve was constructed, and all samples were measured in duplicate. To ensure a low background, a ‘blank’ containing only rL/L reagent and HBSS was analyzed. ATP concentrations were determined by comparison to a standard curve.

Shu T., Lv Z., Xie Y., Tang J, & Mao X. (2019). Hepcidin as a key iron regulator mediates glucotoxicity-induced pancreatic β-cell dysfunction. Endocrine Connections, 8(3), 150-161.

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Dual Luciferase Activity Assay Protocol

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For dual luciferase activity assay, each reporter construct was co-transfected with Renila Luciferase plasmid for three biological replicates. Briefly, plasmids were electroporated into GMSM-K cells (1 × 10⁶ per transfection) with Amaxa Cell Line Nucleofector Kit V (Lonza, Cologne, Germany) using Nucleofector II (Lonza) (program: X-005), and plasmids were electroporated into HEPM cells (1 × 10⁶ per transfection) with Amaxa Basic Nucleofector Kit for Primary Mammalian Fibroblasts (Lonza) using Nucleofector II (Lonza) (program: U-020). The Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA) and 20/20n Luminometer (Turner Biosystems, Sunnyvale, CA, USA) were employed to evaluate the luciferase activity when cells were approximately 95% confluent at 72 h post-transfection following the manufacturer's instructions. Relative luciferase activities were calculated by the ratio between the value for firefly and Renilla luciferase activities. Three measurements were made for the lysate from each transfection group. All quantified results are presented as mean±s.d. Student t-test was used to determine statistical significance.

Liu H., Leslie E.J., Carlson J.C., Beaty T.H., Marazita M.L., Lidral A.C, & Cornell R.A. (2017). Identification of common non-coding variants at 1p22 that are functional for non-syndromic orofacial clefting. Nature Communications, 8, 14759.

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Transcriptional Activity Assay Protocol

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Cells were seeded in 12-well plates at the density of 2.5 × 10⁵ cells per well. After 24 h, pRL-TK (rellina luciferase vector for normalization, 0.75 ng/well) and p3xARE-∆56-c-Fos-GL3 (reporter gene vector) were co-transfected by using PolyJet in vitro DNA transfection reagent (SigmaGen Laboratories) for 5 h, and substituted medium containing DHT or/and CAPE for 48 h. Cell lysates were lysed in 100 μl 1X passive lysis buffer (Promega). Dual-luciferase reporter assay kit (Promega) was used to measure transcriptional activity by Turner Biosystems 20/20n Luminometer.

Kuo Y.Y., Huo C., Lin C.Y., Lin H.P., Liu J.S., Wang W.C., Chang C.R, & Chuu C.P. (2019). Caffeic acid phenethyl ester suppresses androgen receptor signaling and stability via inhibition of phosphorylation on Ser81 and Ser213. Cell Communication and Signaling : CCS, 17, 100.

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Wnt/β-Catenin Activity Quantification

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The Wnt/β-catenin activity was accessed by a dual luciferase reporter assay using a Dual-Glo (TM) Luciferase Assay System (Promega, Madison, WI, USA) on a 20/20n Luminometer (Turner BioSystems, Sunnyvale, CA, USA) essentially according to the manufacturer’s protocols.

Wu X., Deng G., Hao X., Li Y., Zeng J., Ma C., He Y., Liu X, & Wang Y. (2014). A Caspase-Dependent Pathway Is Involved in Wnt/β-Catenin Signaling Promoted Apoptosis in Bacillus Calmette-Guerin Infected RAW264.7 Macrophages. International Journal of Molecular Sciences, 15(3), 5045-5062.

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NFkB-Mediated Gene Expression Quantification

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A luciferase adenoviral expression vector (AdNFkB-Luc) (Vector Biolabs) controlled by a promoter containing five repeats of the NFkB enhancer element (TGGGGACTTTCCGC) was used to infect HCAECs that were simultaneously infected with a β -galactosidase expression vector (Vector Biolabs). HCAEC were infected with adenovirus for 24 h prior to treatments. Cell lysates were obtained using the reporter lysis buffer (Promega) and promptly used for the luciferase and β-galactosidase enzyme assays (Promega). Firefly luciferase activity was measured using the 20/20ⁿ luminometer (Turner Biosystems), while β-galactosidase activity was measured by absorbance detection at 420 nm (SPECTRA MAX 190, Molecular Devices).

Barroso M., Kao D., Blom H.J., de Almeida I.T., Castro R., Loscalzo J, & Handy D.E. (2015). S-Adenosylhomocysteine induces inflammation through NFkB: a possible role for EZH2 in endothelial cell activation. Biochimica et biophysica acta, 1862(1), 82-92.

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Luciferase Assay for Transfected Cell Lines

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All cell lines (U266, CAG, RPMI8226, KG-1, Reh, HT-1080, A549, H1229, and PC-3) were seeded in six-well culture dishes according to Lonza instruction (AMAXA Biosystems, Lonza, Germany). For the luciferase assay, cells were transfected with 2 µg of each DNA contract using the Ingenio Electroporation Kit (Mirus Bio, Madison, WI) and Nucleofector^TM (AMAXA biosystems, Lonza, Germany).
Twenty-four hours after electroporation, the cells were lysed in passive lysis buffer (Promega, Madison, WI). The luciferase assay system (Promega, Madison, WI) and 20/20ⁿ Luminometer (Turner BioSystems, Sunnyvale, CA) were used for the firefly luciferase activity measurement. Two methods were applied for normalization. The first was total protein quantification according to the Bradford method (Bio-Rad, Hercules, CA). pGL4.73 (hRluc/SV40) vector was used as an internal control vector (Promega, Madison, WI) in the second method. Luciferase activity of pGL4.26 without insert was defined as a 100%. The data shown are representative examples of at least three experiments, which were performed in triplicates. Differences in luciferase activities among constructs were determined by t-test. A p value of ≤0.05 was considered as statistically significant.

Ostrovsky O., Grushchenko-Polaq A.H., Beider K., Mayorov M., Canaani J., Shimoni A., Vlodavsky I, & Nagler A. (2018). Identification of strong intron enhancer in the heparanase gene: effect of functional rs4693608 variant on HPSE enhancer activity in hematological and solid malignancies. Oncogenesis, 7(6), 51.

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Dual Luciferase Assay for Gene Expression

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For dual luciferase assays, each reporter construct was co-transfected with Renilla luciferase plasmid and three biological replicates were used. Briefly, GMSM-K cells were electroporated with plasmid using the Amaxa Cell Line Nucleofector Kit (Lonza, Cologne, Germany) and the Nucleofector II instrument (Lonza). We used a dual-luciferase reporter assay system (Promega, Madison, WI) and 20/20 n Luminometer (Turner Biosystems, Sunnyvale, CA) to evaluate the luciferase activity following manufacturer’s instructions. Relative luciferase activity was calculated as the ratio between the value for the Firefly and Renilla enzymes. Three independent measurements were performed for each transfection group. All results are presented as mean ±s.d. Statistical significance was determined using the Student’s t-test.

Liu H., Duncan K., Helverson A., Kumari P., Mumm C., Xiao Y., Carlson J.C., Darbellay F., Visel A., Leslie E., Breheny P., Erives A.J, & Cornell R.A. (2020). Analysis of zebrafish periderm enhancers facilitates identification of a regulatory variant near human KRT8/18. eLife, 9, e51325.

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Evaluating Gene Transfer Efficiency of DNA Nanoparticles

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The in vitro gene transfer efficiency of CH₁₂K₁₈PEG_5k DNA nanoparticles was evaluated using the reporter plasmid, pRNAT-H1.3/Hygro/siFluc (GenScript, Piscataway, NJ). This plasmid has a coral GFP reporter (cGFP) under CMV promoter control, which can be used to track the transfection efficiency; the plasmid also contains the siRNA construct for firefly luciferase, which uses a CMV-enhanced H1 promoter for siRNA expression. GL261 cells were seeded onto 12-well plates at an initial density of 5.0×10⁴ cells per well and incubated overnight at 37°C. After 24 h, cells were incubated with DNA nanoparticles (2 µg DNA/well) in media for 6 hrs at 37°C. Subsequently, nanoparticles and culture medium were replaced with fresh media. After additional 48 h incubation at 37°C, GFP positive cells were sorted and collected by FACS, after which the GFP positive cell populations were imaged under confocal microscope. Subsequently, 0.25 ml of 1X Reporter Lysis Buffer was added to each wells and then subjected to a freeze-and-thaw cycle. Cells were detached and collected using a cell scraper, and supernatants were obtained by centrifugation. Luciferase activity in the supernatants was analyzed using a standard luciferase assay kit (Promega, Madison, WI) and a 20/20n luminometer (Turner Biosystems, Sunnyvale, CA). Data are shown as relative light unit (RLU)/mg protein.

Kim A.J., Woodworth G.F., Boylan N.J., Suk J.S, & Hanes J. (2014). Highly compacted pH-responsive DNA nanoparticles mediate transgene silencing in experimental glioma. Journal of materials chemistry. B, Materials for biology and medicine, 2(46), 8165-8173.

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