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Victor2 luminometer

Manufactured by PerkinElmer
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The Victor2 luminometer is a versatile and reliable laboratory instrument designed for measuring luminescence-based assays. It provides accurate and reproducible results for a wide range of applications, including cell-based assays, reporter gene studies, and biochemical analyses. The Victor2 luminometer offers high sensitivity, a wide dynamic range, and rapid data acquisition, making it a valuable tool for researchers in various fields of study.

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Lab products found in correlation

Dual glo luciferase assay kit, by Promega (2 mentions) Change it multiple mutation site directed mutagenesis kit, by Thermo Fisher Scientific (2 mentions) Celltiter glo luminescent, by Promega (2 mentions) Transcriptaid t7 high yield transcription kit, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Promega (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Dual luciferase reporter assay system, by Promega (1 mentions) Cytofix cytoperm, by BD (1 mentions) Golgistop, by BD (1 mentions) Reporter lysis buffer, by Promega (1 mentions) Passive lysis buffer (plb), by Promega (1 mentions) Rapamycin, by Merck Group (1 mentions) Pgl3 basic expression vector, by Promega (1 mentions) Duoset elisa test, by R&D Systems (1 mentions) Anti cd3 145 2c11, by Exbio (1 mentions)

Spelling variants of this product

Victor2 (111 citations) Wallac Victor2 luminometer (2 citations)

10 protocols using victor2 luminometer

1

Dual-Luciferase Reporter Gene Assay

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Cells in 12-well plates were transfected with firefly and pRL-T7-renilla (Promega) plasmids at a ratio of 100:1 using Lipofectamine LTX as described above. The cells were harvested 48 h later and dispensed equally into a 96-well luminometer plate. Luciferase assay was performed using the Dual-Glo luciferase assay kit (Promega) following the manufacturer’s instruction, and the luminescence was measured in a Victor II luminometer (Perkin Elmer). Firefly luminescence readings were normalised with the corresponding renilla readings. Data presented are mean ± SD from three independent transfection experiments.
Zhang S., Bell E., Zhi H., Brown S., Imran S.A., Azuara V, & Cui W. (2019). OCT4 and PAX6 determine the dual function of SOX2 in human ESCs as a key pluripotent or neural factor. Stem Cell Research & Therapy, 10, 122.
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2

Measuring Amino Acid Variant Effects on HCV Inhibitor Activity

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The methods for measuring the effects of individual amino acid variants on the activity of an inhibitor in HCV replicon cell culture assays were described previously (17 (link)). Variants in NS3, NS5A, or NS5B were each introduced into the GT1a strain H77 replicon using a Change-IT multiple-mutation site-directed mutagenesis kit (Affymetrix, Santa Clara, CA). After the presence of the variant(s) was confirmed by sequence analysis, the plasmid was linearized and a TranscriptAid T7 high-yield transcription kit (Fermentas, Glen Burnie, MD) was used to transcribe the HCV genomic RNA from the plasmid. In a transient-transfection assay, the replicon RNA containing the variant was transfected via electroporation into a Huh-7 cell line (16 (link)). The luciferase activity in the cells was measured using a Victor II luminometer (Perkin-Elmer, Waltham, MA). The 50% effective concentration was calculated using nonlinear regression curve fitting to the 4-parameter logistic equation and GraphPad Prism (version 4) software.
Krishnan P., Tripathi R., Schnell G., Reisch T., Beyer J., Irvin M., Xie W., Larsen L., Cohen D., Podsadecki T., Pilot-Matias T, & Collins C. (2015). Resistance Analysis of Baseline and Treatment-Emergent Variants in Hepatitis C Virus Genotype 1 in the AVIATOR Study with Paritaprevir-Ritonavir, Ombitasvir, and Dasabuvir. Antimicrobial Agents and Chemotherapy, 59(9), 5445-5454.
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3

Dual-Luciferase Assay for hESCs

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hESCs were split 1:6 into 12-well plates using the accutase/ROCKi method as above and allowed to reach 70–80% confluence prior to transfection. Both firefly pGL3-CAGA12-luc and renilla pRL-T7-renilla (Promega) plasmids were transfected at a ratio of 10:1 using Lipofectamine 2000 following the manufacturer's protocol. Forty-two hours post transfection, hESCs were differentiated for 6 h, harvested using accutase and dispensed into a 96-well luminometer plate. Luciferase assay was performed using the Dual-Glo luciferase assay kit (Promega) following the accompanying protocol. Both firefly and renilla luminescence was recorded using a Victor II luminometer (Perkin Elmer). Firefly luminescence readings were normalized with corresponding renilla luminescence readings to give representative luciferase expression values that are independent of transfection efficiency and cell number. Data presented are mean±s.d. from three independent transfection experiments.
Yu J.S., Ramasamy T.S., Murphy N., Holt M.K., Czapiewski R., Wei S.K, & Cui W. (2015). PI3K/mTORC2 regulates TGF-β/Activin signalling by modulating Smad2/3 activity via linker phosphorylation. Nature Communications, 6, 7212.
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4

Measuring Amino Acid Variant Effects on HCV Inhibitor Activity

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The methods describing the measurement of the effects of individual amino acid variants on the activity of an inhibitor in HCV replicon cell culture assays were described previously (11 (link), 12 (link)). NS3 and NS5A variants each were introduced into the GT1b-Con1 subgenomic replicon plasmid using the Change-IT multiple-mutation site-directed mutagenesis kit (Affymetrix, Santa Clara, CA). In a transient assay, the replicon RNA containing the variant was transfected via electroporation into an Huh-7 cell line (31 (link), 32 (link)). The luciferase activity in the cells was measured using a Victor II luminometer (Perkin-Elmer, Waltham, MA). The 50% effective concentrations (EC50s) of paritaprevir and ombitasvir, which were synthesized at AbbVie (33 (link)), were calculated using a nonlinear regression curve fit to the 4-parameter logistic equation in GraphPad Prism 4 software.
Krishnan P., Schnell G., Tripathi R., Beyer J., Reisch T., Zhang X., Setze C., Rodrigues L J.r., Burroughs M., Redman R., Chayama K., Kumada H., Collins C, & Pilot-Matias T. (2016). Analysis of Hepatitis C Virus Genotype 1b Resistance Variants in Japanese Patients Treated with Paritaprevir-Ritonavir and Ombitasvir. Antimicrobial Agents and Chemotherapy, 60(2), 1106-1113.
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5

GRP94 Impact on Wnt Signaling

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To evaluate the activating role of GRP94 on Wnt signaling, GRP94-knockouted or parental MDA-MB231 cells were transfected with the TCF reporter plasmid (TOPFlash) and β-galactosidase plasmid as a control to monitor the transfection efficiency. Transfection was performed with lipofectamine 2000 (Invitrogen) according to the manufacturer’s protocol. After 48 h, the cells were lysed and assayed for luciferase and β-galactosidase activity using Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA), according to the manufacturer’s instructions [59 (link)]. All assays were performed in triplicate using a Victor2 luminometer (Perkin Elmer, Boston, MA).
Kim H.Y., Kim Y.M, & Hong S. (2024). CK2α-mediated phosphorylation of GRP94 facilitates the metastatic cascade in triple-negative breast cancer. Cell Death Discovery, 10, 185.
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6

Proliferation and Cytokine Assay for OT-I CD8+ T Cells

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For proliferation assay, transfected Cas9‐EGFP OT‐I CD8+ T cells were stimulated with various doses of SIINFEKL peptide MHC‐I tetramers (provided by the NIH tetramer core facility) with or without soluble anti‐CD28 (37‐51; EXBIO) antibody. After 48 h of culture, T‐cell proliferation was assessed with CellTiter‐Glo® Luminescent (Promega). The resulting luminescence, which is proportional to the ATP content of the culture, was measured with a Victor 2 luminometer (Wallac, Perkin Elmer Life Science). For IFN‐γ production, cells were stimulated with various doses of N4‐laden pMHC‐I tetramers with soluble anti‐CD28 antibody for 24 h and treated with GolgiStop (BD Biosciences) for the last 4 h. After incubation, cells were washed and stained for dead cells, permeabilized (fixation/permeabilization buffer; Cytofix/CytoPerm BD Biosciences), and stained for intracellular IFN‐γ (XMG1.2; BioLegend) before analysis by flow cytometry.
Locard‐Paulet M., Voisinne G., Froment C., Goncalves Menoita M., Ounoughene Y., Girard L., Gregoire C., Mori D., Martinez M., Luche H., Garin J., Malissen M., Burlet‐Schiltz O., Malissen B., Gonzalez de Peredo A, & Roncagalli R. (2020). LymphoAtlas: a dynamic and integrated phosphoproteomic resource of TCR signaling in primary T cells reveals ITSN2 as a regulator of effector functions. Molecular Systems Biology, 16(7), e9524.
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7

Caspase Activity Assay Protocol

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Caspase-3 activity and caspase-9 activity were measured using the luminescent substrates Z-DEVD-aminoluciferin (Caspase-Glo 3/7 assay; Promega Corporation, Fitchburg, WI, USA) and Z-LEHD-aminoluciferin (Caspase-Glo 9 assay; Promega Corporation), respectively. Then, the experiment was conducted according to the manufacturer’s instructions. Briefly, cells were cultured in DMEM containing 10% FBS. Then, cells were harvested and lysed in the Caspase-Glo reagent provided in the kit, and the luminescence (substrate turnover) of triplicate samples was determined at times 0 and 1 hour using a Victor2 luminometer (PerkinElmer Inc., Waltham, MA, USA). Protein quantification was carried out using the Bradford method. Caspase activities were calculated as Δ luminescence units per microgram protein.
Wang D., Cheng Z., Zhao M., Jiao C., Meng Q., Pan H., Xie Y., Li L., Zhu Y., Wang W., Qu C, & Liang D. (2019). PTPN9 induces cell apoptosis by mitigating the activation of Stat3 and acts as a tumor suppressor in colorectal cancer. Cancer Management and Research, 11, 1309-1319.
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8

Tcf/Lef-Luc Reporter Assay for DHA Treatment

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Cultured cells were seeded at a concentration achieving 80% confluence in 12-well plates for 24 h prior to transfection. The cells were transiently transfected with 0.2 μg/well of a translucent Tcf/Lef-Luc reporter vector, which was designed to measure the transcriptional activity of Tcf/Lef-responsive genes. After transfection, the cells were treated with DHA in serum-free medium at the indicated time periods. The cell lysates were then obtained with 1 × reporter lysis buffer (Promega, Madison, WI). The luciferase activity was assayed using a Victor2 luminometer (Perkin Elmer, Waltham, MA). The relative luciferase activity was calculated after normalization of cellular proteins. All values are expressed as the percentage of activity relative to basal activity.
Han Y.M., Jeong M., Park J.M., Kim M.Y., Go E.J., Cha J.Y., Kim K.J, & Hahm K.B. (2016). The ω-3 polyunsaturated fatty acids prevented colitis-associated carcinogenesis through blocking dissociation of β-catenin complex, inhibiting COX-2 through repressing NF-κB, and inducing 15-prostaglandin dehydrogenase. Oncotarget, 7(39), 63583-63595.
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9

DHPR-α1.1 Promoter Regulation in C2C12 Myoblasts

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The constructs containing DHPRα 1.1 promoter region fragments (Luc7P–724, Luc/P756) were cloned in frame in pGL3-basic expression vector (Promega) [35 (link)]. C2C12 myoblasts (2 × 104 cells per dish) were plated on gelatin-coated 35 mm cell culture dishes and allowed to grow in DMEM plus Glutamax, 4.5 g/l glucose, 20% FBS, 50 units/ml penicillin, 50 μg/ml streptomycin until 50% confluent. Two microgram of each construct and 200 ng of the control vector pRL–TK were used for transfection using the FuGENE6 transfection reagent (Roche). Twenty-four hours after transfection, cells were induced to differentiate by changing the medium to DMEM plus Glutamax, 4.5 g/l glucose, 5% horse serum and penicillin/streptomycin. After 4–6 days, when cells had visibly fused into mytoubes, 20 μM rapamycin (Sigma) was added as described [36 (link),37 (link)]. Cells were then washed twice with PBS and lysed using Passive Lysis Buffer (Promega). Firefly luciferase and renilla activity were measured with Victor2 Luminometer (PerkinElmer). Firefly luciferase activity was normalized to renilla activity and expressed as mean (± S.E.M.) enzymatic activity units.
Lopez R.J., Mosca B., Treves S., Maj M., Bergamelli L., Calderon J.C., Bentzinger C.F., Romanino K., Hall M.N., Rüegg M.A., Delbono O., Caputo C, & Zorzato F. (2015). Raptor ablation in skeletal muscle decreases Cav1.1 expression and affects the function of the excitation–contraction coupling supramolecular complex. The Biochemical journal, 466(1), 123-135.
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10

T Cell Proliferation and IL-2 Assay

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For proliferation and IL‐2 secretion assay, purified CD4+ T cells were stimulated with plate‐bound anti‐CD3 (145‐2C11; Exbio) and soluble anti‐CD28 (37–51; Exbio). After 48 h of culture, T‐cell proliferation was assessed with CellTiter‐Glo® Luminescent (Promega). The resulting luminescence, which is proportional to the ATP content of the culture, was measured with a Victor 2 luminometer (Wallac, Perkin Elmer Life Science). IL‐2 production was measured with a DuoSet ELISA test (R&D Systems).
Voisinne G., García‐Blesa A., Chaoui K., Fiore F., Bergot E., Girard L., Malissen M., Burlet‐Schiltz O., Gonzalez de Peredo A., Malissen B, & Roncagalli R. (2016). Co‐recruitment analysis of the CBL and CBLB signalosomes in primary T cells identifies CD5 as a key regulator of TCR‐induced ubiquitylation. Molecular Systems Biology, 12(7), 876.
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