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Sirius single tube luminometer

Manufactured by Berthold Technologies
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Sourced in Germany

The Sirius Single Tube Luminometer is a compact and versatile instrument designed for the detection and measurement of luminescence signals. It utilizes a single detection tube to provide accurate and reliable luminescence measurements, making it a suitable tool for a variety of applications.

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Lab products found in correlation

Dual luciferase reporter assay system, by Promega (1 mentions) Transit lt1, by Mirus Bio (1 mentions) Galacto light plus kit, by Thermo Fisher Scientific (1 mentions) Dlr assay system, by Promega (1 mentions) Lipofectamine ltx, by Thermo Fisher Scientific (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Dual luciferase reporter assay system kit, by Promega (1 mentions) Reporter lysis buffer, by Promega (1 mentions) Dual glo luciferase assay system, by Promega (1 mentions) N decyl aldehyde, by Merck Group (1 mentions) Dual luciferase assay reagent, by Promega (1 mentions) Nano glo assay kit, by Promega (1 mentions) Infinite m200 plate reader, by Tecan (1 mentions) 6 well plate, by Corning (1 mentions)

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Sirius luminometer (55 citations) Tube luminometer (14 citations) Single Tube Luminometer (4 citations) Sirius L single tube luminometer (3 citations) Sirius tube luminometer (2 citations)

9 protocols using sirius single tube luminometer

1

Validating miR-1956 Target Genes via Luciferase Assay

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To validate miR-1956 target genes, we performed luciferase reporter assays on HEK293 cells as previously described 21 (link). Briefly, cells were transiently transfected using TransIT-LT1 (Mirus) according to the manufacturer's protocol. A dual-luciferase reporter assay system (Promega) and a Sirius single tube luminometer (Berthold) were used to determine luciferase activity. The Renilla luciferase coding vector (Promega) was co-transfected in all cases to evaluate transfection efficiency. Luciferase experiments were performed in quadruplicate and repeated three times independently. Luciferase activity results are presented as relative light units (RLU): the average of Photinus pyralis luciferase activity was divided by the average of Renilla luciferase activity.
Gao L., Mei S., Zhang S., Qin Q., Li H., Liao Y., Fan H., Liu Z, & Zhu H. (2020). Cardio-renal Exosomes in Myocardial Infarction Serum Regulate Proangiogenic Paracrine Signaling in Adipose Mesenchymal Stem Cells. Theranostics, 10(3), 1060-1073.
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2

Luciferase Assay Protocol for siRNA Experiments

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For eS25-siRNA experiments, luciferase assays were performed on 4 μl of lysate using the DLR™ Assay System (#E1960; Promega), and β-galactosidase activity was measured using 6 μl of lysate with the Galacto-Light Plus kit (ThermoFisher Scientific) according to the manufacturers’ protocols with a Lumat LB 9507 luminometer (Berthold Detection Systems GmbH). For all other luciferase assays 20 μl of cell lysate or 1 μl of RRL reaction was measured using a Sirius Single Tube Luminometer (Berthold). The activity of firefly luciferase (FLuc) and Renilla luciferase (RLuc) were measured using the DLR™ Assay System according to manufacturer’s instructions.
Cáceres C.J., Angulo J., Lowy F., Contreras N., Walters B., Olivares E., Allouche D., Merviel A., Pino K., Sargueil B., Thompson S.R, & López-Lastra M. (2018). Non-canonical translation initiation of the spliced mRNA encoding the human T-cell leukemia virus type 1 basic leucine zipper protein. Nucleic Acids Research, 46(20), 11030-11047.
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3

Protein Extraction and Quantification for Western Blot

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Cell lysates were prepared 24 h post-transfection in lysis buffer C (50 mM Tris–HCl pH 7.8, 100 mM NaCl, 0.5% IGEPAL). The concentration of total protein in the lysate was determined by Bradford assay. Equal amounts of protein were loaded in SDS-PAGE and processed for western blotting.
Luciferase activity (RLU) was determined using Berthold Sirius Single Tube Luminometer, and then normalized to the amount of protein in the lysate (RLU/µg). Each experiment was repeated independently at least three times. Values represent the mean ± SEM (standard error of the mean).
Embarc-Buh A., Francisco-Velilla R., Garcia-Martin J.A., Abellan S., Ramajo J, & Martinez-Salas E. (2022). Gemin5-dependent RNA association with polysomes enables selective translation of ribosomal and histone mRNAs. Cellular and Molecular Life Sciences, 79(9), 490.
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4

Bicistronic Construct Transfection Assay

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HEK293 cell monolayers at 80 % of confluency were co-transfected with the bicistronic construct CMVpBIC and pcDNA3-Xpress-G5845-1508 WT or the different mutants using Lipofectamine LTX (Thermo Fisher Scientific) and were harvested 24 hpt. Cell lysates were prepared in lysis buffer C (50 mM Tris-HCl pH 7.8, 100 mM NaCl, 0.5 % IGEPAL). The concentration of total protein in the lysate was determined by the Bradford assay (BioRad). IRES activity was quantified as the expression of Luciferase activity (RLU) using Berthold Sirius Single Tube Luminometer and normalized to the amount of protein in the lysate. CAT activity, which reported cap-dependent translation, was determined by the liquid scintillation assay [30] (link). Appropriate dilutions of the extracts were chosen to be in the linear range of the assay. Values represent the mean ± SEM (standard error of the mean). Differences in distribution between two samples were analyzed by paired two-tailed Student’s t-test and were considered significant when P < 0.05. The resulting P-values were graphically illustrated in figures with asterisks as described in figure legends.
Francisco-Velilla R., Embarc-Buh A., Abellan S., del Caño-Ochoa F., Ramón-Maiques S, & Martinez-Salas E. (2022). Phosphorylation of T897 in the dimerization domain of Gemin5 modulates protein interactions and translation regulation. Computational and Structural Biotechnology Journal, 20, 6182-6191.
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5

Dual-Luciferase Assay of pGL3-Enhancer Constructs

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The different pGL3-enhancer constructs were transfected into HEK293T and hTERT-RPE1 using FuGENE-HD® Transfection Reagent as previously indicated. The pRL-CMV Renilla luciferase reported plasmid (Promega) was co-transfected for the internal control of cell transfection using the Dual-Luciferase reporter assay system kit (Promega), according to the manufacturer’s instructions. Firefly and Renilla luminescence were measured 48 h after the transfection using a luminometer (Berthold Sirius Single Tube Luminometer, Berthold Technologies (Bad Wildbad, Germany)). The Luciferase measurement was performed in duplicate using 5 µL of the cell lysate. The fold-change induction was estimated as the ratio of the Firefly between the Renilla average values. The data were normalized against the XS construct.
Vázquez-Domínguez I., Duijkers L., Fadaie Z., Alaerds E.C., Post M.A., van Oosten E.M., O’Gorman L., Kwint M., Koolen L., Hoogendoorn A.D., Kroes H.Y., Gilissen C., Cremers F.P., Collin R.W., Roosing S, & Garanto A. (2022). The Predicted Splicing Variant c.11+5G>A in RPE65 Leads to a Reduction in mRNA Expression in a Cell-Specific Manner. Cells, 11(22), 3640.
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6

Quantifying APP Intracellular Domain Release

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SH-SY5Y cells were co-transfected with Lipo2000® in a 1:1:1 ratio with pG5E1B-luc, pRL-TK, and either a pCDNA3.1-empty plasmid (EP) or the pCDNA3.1-C99-GVP, bearing a tagged C99 to quantify the release of the APP intracellular domain (AICD). The system setup was previously described [21 (link), 32 (link)]. Cells were rinsed with PBS 48 h after transfection and incubated with the reporter lysis buffer (Promega, Madison, WI, USA) for 15 min at rt. Firefly and Renilla luciferase activities were measured using the Dual-Glo® luciferase assay system (Promega, Madison, WI, USA) on a Sirius single tube luminometer (Berthold, Bad Wildbad, Germany). Luciferase activity corrected for transfection efficiency was calculated as the Firefly/Renilla ratio.
Vrancx C., Vadukul D.M., Suelves N., Contino S., D’Auria L., Perrin F., van Pesch V., Hanseeuw B., Quinton L, & Kienlen-Campard P. (2021). Mechanism of Cellular Formation and In Vivo Seeding Effects of Hexameric β-Amyloid Assemblies. Molecular Neurobiology, 58(12), 6647-6669.
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7

Luminometric Bacterial Viability Assay

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Bacterial luciferase (Vibrio harveyi) previously introduced into Mycobacterium smegmatis mc2155 with the vector pSMT3LxEGFP, was provided by Brian D. Robertson (Imperial College London) [30 (link)]. Luminescence measurements were obtained using a Berthold Sirius single tube luminometer (Berthold Technologies, Bad Wildbad, Germany). Ten μl samples were diluted with 190 μl PBS, pH 7.4. Bacterial luminescence was measured immediately after addition of 20 μl substrate, 1% n-decyl aldehyde (Sigma-Aldrich, St. Louis, MO, USA) in ethanol, for 10 s using an integration time of 1 s, and the results were expressed in relative light units (RLU). Sterile PBS without bacteria was used to measure background and subtracted from sample RLU measurements. The RLU:CFU ratio was determined to be 10:1. The exponential growth rate constant (k) was determined under control conditions using k=ln(N/ N0)/t-to where N0= initial number of bacteria at initial time to, and N= number at time t. The percent survival of cells was determined in the experimental groups from the ratio of the number of bacteria observed from that expected at 48 hours given k above and N0 verified from the RLU at time 0 for each experiment.
Staudinger T., Redl B, & Glasgow B.J. (2014). Antibacterial Activity of Rifamycins for M. Smegmatis with Comparison of Oxidation and Binding to Tear Lipocalin. Biochimica et biophysica acta, 1844(4), 750-758.
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8

Luciferase Activity Quantification

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Quantification of luciferase activities were performed with commercially available kits (Promega, Mannheim, Germany) in concordance to the manufacturer’s instructions. Frozen cells from sub-experiments (i), (ii), (iii) and (iv) were incubated with the passive lysis buffer enclosed in the respective luciferase assay kit for 20 min at room temperature. Lysates from (i) and (ii) were combined with dual luciferase assay reagents (Promega, #E1910), whereas lysates from (iii) and (iv) were mixed with the Renilla assay reagent (Promega, #E2810). Bioluminescence was measured in a Sirius single-tube luminometer (Berthold Technologies, Bad Wildbad, Germany). In (i) and (ii) PLuc activities were normalized to those of the constitutively expressed RLuc enzyme derived from phRG-b vector. Medium and cells collected in experiment (v) were processed with compounds from the Nano Glo assay kit (Promega, #N1110) in accordance to the manufacturer’s instructions. secNLuc bioluminescence was quantified in an Infinite M200 plate reader (Tecan, Männedorf, Switzerland).
Bolze F., Mocek S., Zimmermann A, & Klingenspor M. (2017). Aminoglycosides, but not PTC124 (Ataluren), rescue nonsense mutations in the leptin receptor and in luciferase reporter genes. Scientific Reports, 7, 1020.
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9

Transcriptional Activation of p21 Promoter

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For transcriptional activation analysis of the p21 promoter, H1299 cells were transiently transfected with different amounts of p53, MDM2 and ZNF217, p21luciferase and load control RT-luciferase plasmid constructs in 6-well plates (Corning) for 18 h, using the calcium phosphate method. They were then harvested and luciferase activity was measured according to the manufacturer's instructions (Promega, USA) in a Berthold luminometer (Berthold Sirius Single Tube Luminometer, Germany).
Mantsou A., Koutsogiannouli E., Haitoglou C., Papavassiliou A.G, & Papanikolaou N.A. (2016). Regulation of expression of the p21CIP1 gene by the transcription factor ZNF217 and MDM2. Biochemistry and cell biology = Biochimie et biologie cellulaire, 94(6).
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