Synergy ht luminometer

Manufactured by Agilent Technologies

Sourced in United States

The Synergy HT luminometer is a highly sensitive and versatile instrument designed for a wide range of luminescence-based assays. It features a high-performance photomultiplier tube (PMT) detector and advanced optics to provide accurate and reproducible measurements. The Synergy HT is capable of detecting a broad range of luminescent signals, making it suitable for a variety of applications in life science research and drug discovery.

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Lab products found in correlation

Caspase glo 3 7, by Promega (3 mentions) Dual luciferase reporter assay system kit, by Promega (2 mentions) Celltiter glo luminescent cell viability assay kit, by Promega (2 mentions) Luciferase assay kit, by Promega (2 mentions) Passive lysis buffer (plb), by Promega (2 mentions) Dual luciferase reporter assay system, by Promega (2 mentions) Pcmv6 xl6 plasmid, by OriGene (2 mentions) Originpro 8, by OriginLab (2 mentions) Celltiter glo, by Promega (2 mentions) Attractene transfection reagent, by Qiagen (1 mentions) Data analysis software, by Qiagen (1 mentions) Dual luciferase reporter assay kit, by Promega (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Prl tk plasmid, by Promega (1 mentions) Tnf α, by R&D Systems (1 mentions) Tgf β1, by Promega (1 mentions) Tnf α, by Promega (1 mentions) Tgf β1, by R&D Systems (1 mentions) Dual luciferase reporter assay system kit, by Agilent Technologies (1 mentions) Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Synergy HT (3550 citations) Synergy HT microplate luminometer (2 citations)

24 protocols using synergy ht luminometer

Caspase-3/7 Activity Luminescent Assay

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Caspase-3/7 activity was measured by a luminescent assay, Caspase-Glo^® 3/7 (Promega Corporation, Wisconsin, U.S.), according to a previously published protocol¹⁶. Luminescence was read in a Synergy HT luminometer (BioTek Instruments, Vermont, U.S.). Luminescence produced by the assay was proportional to the amount of caspase activity present in the cell sample. Results were expressed as LCU/min and normalized to a million viable cells.

Bluhme E., Henckel E., Gramignoli R., Kjellin T., Hammarstedt C., Nowak G., Karadagi A., Johansson H., Jynge Ö., Söderström M., Fischler B., Strom S., Ellis E., Hallberg B, & Jorns C. (2022). Procurement and Evaluation of Hepatocytes for Transplantation From Neonatal Donors After Circulatory Death. Cell Transplantation, 31, 09636897211069900.

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Cignal Finder Assay for Pathway Analysis

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Cignal Finder Reporter Array consists of 45 dual-luciferase reporter assays enabling rapid and reliable identification of signaling pathway activities and to determine the effects of various treatments. Cell-based Multi-Pathway Activity Assay was performed in A2780 cell line (6 × 10⁴ cells/well) according to instructions of the manufacturer. Transfection was performed using Attractene Transfection Reagent (0.3 μl/well, Qiagen). Twenty four hours post transfection, medium was replaced with fresh one, containing SFN (20 μM), Xest (1 μM), combination of both or DMSO only, which served as a controland cells were treated for additional 24 hours. The activity of each signaling pathway was analyzed using Dual Luciferase Reporter Assay Kit (Promega, USA) and measured using Biotek Synergy HT luminometer. Results were calculated for each transfectant and analyzed by the Data Analysis Software (SABiosciences, USA). The change in the activity of each signaling pathway was determined by comparing the normalized luciferase activities of the reporter in SFN/Xest treated versus SFN treated transfectants.

Hudecova S., Markova J., Simko V., Csaderova L., Stracina T., Sirova M., Fojtu M., Svastova E., Gronesova P., Pastorek M., Novakova M., Cholujova D., Kopacek J., Pastorekova S., Sedlak J, & Krizanova O. (2016). Sulforaphane-induced apoptosis involves the type 1 IP3 receptor. Oncotarget, 7(38), 61403-61418.

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Assessing NF-κB Transcriptional Activity

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For testing the NF-κB transcriptional activity, the 1 μg pGL6-NF-κB-Luc plasmid (Beyotime Institute Biotechnology, China) and 1 μg pRL-TK plasmid (Promega) as internal control were transiently co-transfected into LX-2 cells (1 × 10⁵ cells/well) using Lipofectamine 3000 (Invitrogen). After transfection for 12 h, cells were pre-treated with 10 μM NPLC0393, 10 μM SB431542, or 50 ng/mL NF-κB inhibitor SN50 for 12 h and were stimulated with 5 ng/mL TGF-β1 or 25 ng/mL TNF-α (R&D Systems, USA) for another 24 h.
For the measurement of MAT2A promoter activity, the Human MAT2A promoter-luciferase reporter plasmid containing binding sites for NF-κB (Sangon, China) and pRL-TK were transfected into LX-2 cells. After 12 h, cells were pre-treated with 10 μM NPLC0393 or 50 ng/mL SN50, a specific NF-κB translocation inhibitor, for 12 h and were stimulated with 5 ng/mL TGF-β1 or 25 ng/mL TNF-α for another 24 h. Dual luciferase assays were carried out according to the manufacturer's protocol (Promega) and light intensity was measured in a Synergy HT luminometer (BioTek, US). Each experiment was done in triplicate samples and results were normalized against those of cotransfected pRL-TK.

Wang K., Fang S., Liu Q., Gao J., Wang X., Zhu H., Zhu Z., Ji F., Wu J., Ma Y., Hu L., Shen X., Gao D., Zhu J., Liu P, & Zhou H. (2019). TGF-β1/p65/MAT2A pathway regulates liver fibrogenesis via intracellular SAM. EBioMedicine, 42, 458-469.

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Transcription Factor Activation Assay

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HEK293T cells were cultured in Dulbecco’s Modified Eagle’s Medium supplemented with 10% fetal bovine serum and 1% Penicillin/streptomycin, at 37°C and 5% CO₂. For transfection, HEK293T cells were plated in 24-well plates 18–24 h before transfection, at a concentration of 3 × 10⁵ cells/well. Transfection was performed in triplicate and with simultaneous insertion of three different plasmids: one containing the gene coding for the tested transcription factor (Ascl1 and/or Pou3f2), the pGL3-basic which encodes the firefly luciferase under cE1 control and pRL, the vector which produces Renilla luciferase that was used as a normalization factor. Empty pGL3-basic vector was used as control. Co-transfection was performed with 3.3 μL/well Polyethylenimine (PEI), 500 ng/well of transcription factor, 10 ng/well pRL, and 500 ng/well pGL3-cE1 overnight. Each sample was lysed in 1 × lysis buffer from Dual-Luciferase Reporter Assay System kit (Promega, cat. #E1910) and luciferase activity detection was performed according to manufacturer’s instructions in a Synergy HT luminometer (Biotek, United States).

Goes C.P., Botezelli V.S., De La Cruz S.M., Cruz M.C., Azambuja A.P., Simoes-Costa M, & Yan C.Y. (2024). ASCL1 promotes Scrt2 expression in the neural tube. Frontiers in Cell and Developmental Biology, 12, 1324584.

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Luminescent Assay for Cell Viability

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The CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega Italia s.r.l, Milano, Italy) was used to evaluate the ATP levels. For the execution of the assay, 2.5 × 10⁴ cells were seeded in 96-well plates. After equilibrating the plate and the cells at room temperature for about 30 min, 100 µL of CellTiter-Glo® Reagent were added per well. After leaving the plate in agitation for ~2 min to allow cell lysis, the plate was left at room temperature for 10 min to stabilize the luminescent signal, which was subsequently evaluated by BioTek’s Synergy™ HT Luminometer (BioTek Instruments Inc., Winooski, VT, USA).

Pagano C., Navarra G., Coppola L., Avilia G., Pastorino O., Della Monica R., Buonaiuto M., Torelli G., Caiazzo P., Bifulco M, & Laezza C. (2022). N6-isopentenyladenosine induces cell death through necroptosis in human glioblastoma cells. Cell Death Discovery, 8, 173.

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CAR T Cell Cytotoxicity Assay

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Cytotoxic activity of CAR T cells against targets was determined by a luciferase-based cytotoxicity assay. MDA-MB-468, MDA-HER2, JIMT-1, and N87 cells expressing eGFP/ffLuc were plated in 96-well flat-bottom plates at a density of 10⁵ cells/well in triplicates. After 4 h, OKT3-antiCD28/RPMI- or OKT3-RetroNectin/LymphoONE-stimulated effector CAR T cells were added at a 1:1 effector to tumor cell ratio. Wells without effector cells served as untreated controls. After 24 h, luciferase activity was determined using a luciferase assay kit according to the manufacturer’s instructions (Promega, Madison, WI, USA) and a Synergy HT luminometer (BioTek, Winooski, VE, USA). Empty media, NT T cells, HER2+ target cells cultured without the presence of effector cells, and the HER2– MDA-MB-468.ffLuc cell line served as controls.

Csaplár M., Szöllősi J., Gottschalk S., Vereb G, & Szöőr Á. (2021). Cytolytic Activity of CAR T Cells and Maintenance of Their CD4+ Subset Is Critical for Optimal Antitumor Activity in Preclinical Solid Tumor Models. Cancers, 13(17), 4301.

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Evaluating miRNA Regulation of UGT2A1

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Twenty-four hours before transfection, HEK293 cells were seeded onto 24-well plates at a density of approximately 5 × 10⁴ cells/well. The pGL3 plasmids containing either the wild-type or miRNA seed deletion UGT2A1 3′-UTR sequences (380 ng of each) were individually cotransfected with pRL-TK plasmid (20 ng, as a transfection efficiency control) along with 100 nM of miR-196a-5p mimic, miR-196b-5p mimic, or scrambled miRNA control using Lipofectamine 2000 transfection reagent. The HEK293 cells were harvested 24 hours after transfection using passive lysis buffer provided by the DLR kit. We measured the bioluminescent luciferase activity with a Bio-tek Synergy HT luminometer (Winooski, VT) using the Dual-Luciferase Reporter Assay System kit.

Sutliff A.K., Watson C.J., Chen G, & Lazarus P. (2019). Regulation of UGT2A1 by miR-196a-5p and miR-196b-5p. The Journal of Pharmacology and Experimental Therapeutics, 369(2), 234-243.

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Evaluating ERBB2 Reporter Activity

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Twenty-four hours prior to transfections, cells were plated in 96-well plates. Cells were transfected with the specific ERBB2 luciferase reporter construct or the backbone vector (100 ng) plus the internal control plasmid pRL-TK (10 ng) using the Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific). In co-transfection assays, cells were transfected with 100 ng of ERBB2 reporter constructs, 10 ng of pRL-TK, and 150 ng of Sp1 expression vector or empty pCMV6-XL6 plasmid (OriGene, Rockville, MD). Twenty-four hours post transfection, cultures were washed once with PBS; cells were lysed in freshly diluted passive lysis buffer (100 μl/well, Promega) by incubating the plates at room temperature on a shaker at 200 rpm for 60 min. Luciferase reporter gene activities were determined with the Dual-Luciferase Reporter Assay System (Promega) as per manufacturer’s instructions. Light intensity was measured in a Synergy HT luminometer equipped with proprietary software for data analysis (BioTek, Winooski, VT). Corrected firefly luciferase activities were normalized to renilla luciferase activities and expressed relative to the averaged activity of the ₋₄₉₉ERBB2 construct which was assigned an arbitrary value of 100.

Quiñones-Lombraña A., Blair R.H, & Blanco J.G. (2017). Investigation of the Role of DNA Methylation in the Expression of ERBB2 in Human Myocardium. Gene, 628, 286-294.

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MTT Assay for Mithramycin A Cytotoxicity

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The viability of mithramycin A treated A549 cells was assessed by recording the reduction of 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT, Sigma-Aldrich). Briefly, cells were plated in 96 well plates and treated with mithramycin A at different concentrations for 24 h. Then, 20 μl of MTT solution (5 mg/ml) were added to each well followed by incubation at 37 °C for 4 h. After incubation, the medium was removed and 100 μl of DMSO were added into each well. The plate was gently rotated on an orbital shaker for 10 min to dissolve the MTT precipitate. The absorbance at 570 nm was recorded in a Synergy HT luminometer (BioTek). Cellular viability was expressed as percentages relative to control incubations.

Quiñones-Lombraña A., Blair R.H, & Blanco J.G. (2017). Investigation of the Role of DNA Methylation in the Expression of ERBB2 in Human Myocardium. Gene, 628, 286-294.

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Quantifying β-arrestin2 Recruitment to CB2

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For β-arrestin2 enzyme fragment complementation, the PathHunter® β-arrestin assay was performed following the manufacturer’s protocol (DiscoveRx). Briefly, U2OS-βarrestin2-EFC-hCB2 cells were plated at 5000 cells per well in Bio-one CELLSTAR 384 well white plates (Greiner) in AssayComplete™ Cell Plating 2 Reagent (DiscoveRx) overnight at 37°C. The next day, cells were treated with compounds at the indicated doses (prepared as described for cAMP assay) at concentrations ranging from 0.03 – 10,000 nM for 90 min at 37°C followed by 1 hr incubation of PathHunter® detection reagent at room temperature protected from light. Luminescence levels were determined by using a Synergy HT luminometer (BioTek, Winooski, VT) (Hua et al., 2016 (link)). All data points are normalized to vehicle only controls (1% DMSO) included in the assay; all studies were performed with CP55,940 curves run in parallel to assure cellular performance.

Li X., Hua T., Vemuri K., Ho J.H., Wu Y., Wu L., Popov P., Benchama O., Zvonok N., Locke K., Qu L., Han G.W., Iyer M.R., Cinar R., Coffey N.J., Wang J., Wu M., Katritch V., Zhao S., Kunos G., Bohn L.M., Makriyannis A., Stevens R.C, & Liu Z.J. (2019). Crystal Structure of the Human Cannabinoid Receptor CB2. Cell, 176(3), 459-467.e13.

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