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Citations Search

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30, 940-952.  Leveraging an NQO1 bioactivatable drug for tumor-selective use of poly (ADP-ribose) polymerase inhibitors.  2016

Huang, X., Motea, E.A., Moore, Z.R., Yao, J., Dong, Y., Chakrabarti, G., Kilgore, J.A., Silvers, M.A., Patidar, P.L., Cholka, A. and Fattah, F.

Notes: NAD/NADH-Glo™ Assay was used to assess metabolic activity in cells. (4897)

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Cell Biochem. Funct. 34(7), 497-504.
SOD1 dimerization monitoring using a novel split NanoLuc, NanoBit.
2016

Oh-Hashi K., Hirata Y., and Kiuchi K.

Notes: These authors used a NanoBiT® complementation assay to investigate interactions between proteins implicated in development of amyotrophic lateral sclerosis (ALS). Mutations in superoxide dismutase 1 (SOD1) and in TAR‐binding protein 43 kDa (TDP43) have been associated with ALS. The authors observed formation of SOD1 homodimers in neuronal cells transfected with SOD1 tagged with NanoBiT complementation partners, and found that SOD1 mutants associated with ALS were unable to form homodimers.  Next, possible interactions between SOD1 and TDP43 were assessed. When NanoBiT-tagged constructs of TBP43 and SOD were transfected into Neuro 2A cells, only NanoBiT-tagged SOD generated a luciferase signal. TBP43 did not self-associate to form homodimers, and SOD1 and TBP43 did not interact with each other to form heterodimers. The authors state that NanoBiT is a useful and sensitive tool for analyzing protein interactions under physiological conditions, and may provide a convenient way to monitor the modulation of SOD1 conformation by candidate treatments. (4765)

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Sci. Rep. 6, 21548. µ opiod receptor: novel antagonists and structural modeling. 2016

Kaserer, T., Lantero, A., Schmidhammer, H., Spetea, M. and Schuster, D.

Notes: CHO cells expressing the human MOR were stably transfected with the GloSensor™ -22F cAMP Vector using the ViaFect™ Transfection Reagent. Inhibition of the forskolin-stimulated intracellular cAMP accumulation in the cells through the addition of the GloSensor™ cAMP Reagent (4% vol/vol). Assays were performed in 384-well plates with 5,000 cells/well in 30µl. (4668)

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45, 6048-6077. 1001 lights: luciferins, luciferases, their mechanisms of action and applications in chemical analysis, biology and medicine 2016

Kaskova, Z. M., Tsarkova, A. S. and Yampolsky, I. V. 

Notes: This article referenced the NADP/NADPH-Glo™ Assay when describing products for bioluminescent analysis. (4885)

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Sci. Rep. 6, 19103. 3D tumor spheroid model for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained. 2016

Zanoni, M., Piccinini, F., Arienti, C., Zamagni, A., Santi, S., Polico, R., Bevilacqua, A., and Tesei, A.

Notes: The CellTiter-Glo® 3D Cell Viability Assay, Perfecta3D Assay (3Dbiomatrix, Inc.) and simple Trypan Blue exclusion were assessed for measuring cytotoxicity of A549 cell microspheroids. The authors state, “The best and most reproducible method to determine the viability of large spheroids for both chemical and for physical treatments was the CellTiter-Glo 3D Cell Viability assay…” The CellTiter-Glo® 3D Assay reported a linear relationship for spheroids that were 350–650 microns with some deviation from linear for spheroids with a diameter of 850 microns. The authors note that this could be related to physical interference with such large spheroids as well as the presence of a necrotic core in the large spheroids. The luminescence from the CellTiter-Glo® 3D Assays was measured with a GloMax® Instrument. (4642)

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Anal. Biochem. 505, 45–50. A bioluminescent assay for measuring glucose uptake 2016

Valley, M.P., Karassina, N., Aoyama, N., Carlson, C., Cali, J.J. and Vidugiriene, J.

Notes: The authors of this paper describe the application of a non-radioactive, bioluminescent assay to measure glucose uptake. The assay, now available as the Glucose Uptake-Glo™ Assay, showed similar sensitivity and produced comparable results to isotopic methods and is amenable to HTS. Additionally the authors performed the glucose-uptake assay in multiplex with the RealTime-Glo® MT Cell Viability Assay to obtain more information on the health of the cells. Luminescent assay results were measured using the GloMax® Discover Detection System. (4762)

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Nat. Biotechnol. 34, 760-767. A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo. 2016

Chu, J., Oh, Y., Sens, A., Ataie, N., Dana, H., Macklin, J.J., Laviv, T., Welf, E.S., Dean, K.M., Zhang, F., Kim, B.B., Tang, C.T., Hu, M., Baird, M.A., Davidson, M.W., Kay, M.A., Fiolka, R., Yasuda, R., Kim, D.S., Ng, H.L. and Lin, M.Z.

Notes: The authors characterize a novel orange-red fluorescent protein (CyOFP1), which serves as an efficient acceptor for resonance energy transfer from NanoLuc® luciferase. An optimized fusion protein of CyOFP1 and NanoLuc® (Antares) was demonstrated to function as a highly sensitive bioluminescent reporter in vivo, producing substantially brighter signals from deep tissues than firefly luciferase and other bioluminescent proteins. During fluorometry, 50,000 cells in 10 μL Live Cell Imaging Solution were loaded into glass capillary tubes and gently pelleted by centrifugation for 15 s at 500g. 10 μL of Live Cell Imaging Solution with 20 μM D-luciferin, coelenterazine, ViviRen, or furimazine was then added. For BRET6 and Antares-expressing mice, tail vein injections were performed with 333 nmol (123 μg) of furimazine in 120 μL of 8% glycerol, 12% ethanol, 10% hydroxypropyl-β-cyclodextrin, and 35% polyethylene glycol 400 in water. (4760)

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Nucl. Acids Res. 44, 2348-2361. A short conserved motif in ALYREF directs cap- and EJC-dependent assembly of export complexes on spliced mRNAs 2016

Gromadzka, A.M., Steckelberg, A.-L., Singh, K.K, Hofmann, K. and Gehring, N.H.

Notes: NanoLuc® and Firefly luciferase reporters (pCMV128-NanoLuc and pCI-FireflyLuc) were co-transfected into HeLa cells to provide complementary measures of specific mRNA splicing and export activity. In the presence of splicing activity, firefly luciferase pre-mRNA is spliced in the nucleus, exported and translated resulting in firefly luciferase activity. In contrast, the open reading frame of the Nanoluc luciferase in pCMV128 NanoLuc® was placed between two weak splice sites. If the pre-mRNA is spliced, the NanoLuc coding sequence is removed and no luciferase can be expressed. The NanoLuc® reporter can only be translated when the unspliced pre-mRNA is transported to the cytoplasm. pCMV128 NanoLuc® was generated by inserting the NanoLuc® sequence into the NotI and BamHI sites of pCMV128, and then used for export assays. Cells were harvested 72 h post-transfection using 300 μl Passive Lysis Buffer. Lysates were analyzed using the NanoGlo® Dual-Luciferase® Reporter Assay. (4744)

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J. Mol. Diagn. 19, 255–264. A Targeted High-Throughput Next-Generation Sequencing Panel for Clinical Screening of Mutations, Gene Amplifications, and Fusions in Solid Tumors 2016

Luthra, R., Patel, K.P., Routbort, M.J., Broaddus, R.R., Yau, J., Simien, C., Chen, W., Hatfield, D.Z., Medeiros, L.J. and Singh, R.R.

Notes: In a study of 121 tumors, ReliaPrep Large Volume HT gDNA Isolation System was used for nucleic acid extraction from 24 peripheral blood samples. Sequencing library preparation, quantification and next-generation sequencing was performed using the extracted samples and non-Promega products. (4974)

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Nat. Commun. 7, 13665. Acquired RAS or EGFR mutations and duration of response to EGFR blockade in colorectal cancer. 2016

Van Emburgh, B.O., Arena, S., Siravegna, G., Lazzari, L., Crisafulli, G., Corti, G., Mussolin, B., Baldi, F., Buscarino, M., Bartolini, A., Valtorta, E., Vidal, J., Bellosillo, B., Germano, G., Pietrantonio, F., Ponzetti, A., Albanell, J., Siena, S., Sartore-Bianchi, A., Di Nicolantonio, F., Montagut, C. and Bardelli, A.

Notes: The authors purified ctDNA from 1ml plasma using the Maxwell® RSC ccfDNA Plasma Kit with the Maxwell® RSC Instrument. Cell line mismatch repair deficiency was confirmed by the MSI Analysis System, Version 1.2. Cell line authentication was performed using the Cell ID™ and GenePrint® 10 Systems. DNA from cultured and treated cells were purified with the Wizard® SV and SV 96 Genomic DNA Purification Systems. Cetuximab-treated cells were measured for viability with the CellTiter-Glo® Luminescent Cell Viability Assay. (4777)

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Malaria Journal 15, 232. An ultrasensitive NanoLuc-based luminescence system for monitoring Plasmodium berghei throughout its life cycle. 2016

De Niz, M., Stanway, R.R., Wacker, R., Keller, D. and Heussler, V.T.

Notes: NanoLuc® Luciferase was used to monitor Plasmodium berghei throughout it's life cycle, including detecting single parasites in mosquitos, livers and asexual blood stages. (4922)

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Free Radic Bio Med 99, 20–31. Augmentation of glycolytic metabolism by meclizine is indispensable for protection of dorsal root ganglion neurons from hypoxia-iduced mitochondrial compromise.  2016

Zhuo, M., Gorgun, M.F. and Englander, E.W.

Notes: The levels of NADP+ and NADPH in dorsal root ganglion neurons were followed as an indicator of the pentose phosphate branch of glycolysis. Meclizine helped preserve the NADP+/NADPH ratio in hypoxia but was unable to preserve the ratio in the presence of 2-deoxyglucose. As an indicator of the utilization of NADPH, the GSH/GSSG ratio was monitored in the cells.  Meclizine induced increased GSH production during hypoxia which of course was abrogated by 2-deoxyglucose. The study used the NADP/NADPH-Glo™ and GSH/GSSG-Glo™ Assays. (4849)

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Sci. Rep. 6, 28951. Capturing tumor complexity in vitro: Comparative analysis of 2D and 3D tumor models for drug discovery 2016

Stock, K., Estrada, M.F., Vidic, S., Gjerde, K., Rudisch, A., Santo, V.E., Barbier, M., Blom, S., Arundkar, S.C., Selvam, I., Osswald, A., Stein, Y., Gruenewald, S., Brito, C., van Weerden, W., Rotter, V., Boghaert, E., Oren, M., Sommergruber, W., Chong, Y., de Hoogt, R., and Graeser, R.

Notes: To demonstrate the differences between 2D cultures and 3D cultures of various complexities, models were set up for three pathologies: breast (MCF7), prostate (LNCaP) and lung (H1437) carcinomas. The models consisted of mono- or co-cultures of established cell lines with fibroblast cell lines. Co-culture fibroblasts were tissue-derived cancer-associated cell lines with the exception of breast cancer where no CAF was available. MCF7 cells were co-cultured with human dermal fibroblasts. Models were microspheroids free floating in plate wells, alginate microspheroids in bioreactors, and cells on an extracellular matrix. All cell lines were stably transfected with a fluorescent protein and used fluorescence to monitor cell proliferation. CellTiter-Glo® 3D Cell Viability Assay was used to viability of the mono- and co-cultures of alginate microspheres grown in the bioreactors. The method employed for use was quite intensive: 40 minutes under agitation, pipetting 10 times, followed by 40 more minutes of agitation before reading the luminescence.  (4818)

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J. Mech. Behav. Biomed. Materials 57, 246–59. Characterization, mechanical behavior and in vitro evaluation of a melt-drawn scaffold for esophageal tissue engineering. 2016

Tan, Y.J., Yeong, W.Y., Tan, X., An, J., Chian, K.S. and Leong, K.F.

Notes: The RealTime-Glo™ MT Cell Viability Assay was used to assess the growth of L929 murine fibroblast onto a synthetic scaffold. The flat 4 ×4 mm2 scaffolds were seeded with 5 ×103 cells and monitored for proliferation over 48 hours with periodic readings. Growth on standard tissue culture-treated plates was used as a control. (4698)

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Metab Eng 36, 57–67. Combining metabolic engineering and biocompatible chemistry for high-yield production of homo-diacetyl and homo-(S,S)-2,3-butanediol.  2016

Liu, J., Chan, S.H.J., Brock-Nannestad, T., Chen, J., Lee, S.Y., Solem, C., and Jensen, P.R.

Notes: Mid-exponential growth phase Lactococcus lactis cultures (OD600=0.6) were quenched in liquid nitrogen then stored at –20°C until assay. Extraction and quantification of NADH and NAD+ were performed with the NAD/NADH-Glo™ Assay. (4841)

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Cancer Lett. 375, 274-83. CSIRT1 promotes epithelial-mesenchymal transition and metastasis in colorectal cancer by regulating Fra-1 expression. 2016

Cheng, F., Su, L., Yao, C., Liu, L., Shen, J., Liu, C., Chen, X., Luo, Y., Jiang, L., Shan, J., Chen, J., Zhu, W., Shao, J. and Qian, C.

Notes: The ViaFect™ Transfection Reagent was used to transfect Firefly experimental and Renilla control vectors into SW620 to judge the effects and confirm the location of a shRNA to SIRT 1 (introduced through lentiviral vectors) on the SIRT1 promoter. Reporter activity was monitored with the Dual-Luciferase® Reporter Assay System. No details of the transfection were provided. (4656)

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Cancer Biol Ther 17, 1197–205. Dephosphorylation of the retinoblastoma protein (Rb) inhibits cancer cell EMT via Zeb 2016

Egger, J.V., Lane, M.V., Antonucci, L.A., Dedi, B., and Krucher, N.A. 

Notes: The CellTiter-Glo® 3D Cell Viability Assay was used to measure viability of MDA-MB-468 cells grown on Matrigel with inducible knockdown of PNUTS. The effect of PNUTS knockdown was also assessed in a reporter model of E-Cadherin expression using a firefly luciferase reporter vector and Renilla luciferase control vector. Luciferase activities were measured with the Dual-Luciferase® Reporter Assay System. (4823)

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Anal. Chem. 88, 11476–85. Detection and activity profiling of synthetic cannabinoids and their metabolites with a newly developed bioassay. 2016

Cannaert, A., Storme, J., Franz, F., Auwärter, V. and Stove, C.P.

Notes: The authors used NanoLuc® Binary Technology (NanoBiT®) to build β-arrestin recruitment assays for two G-protein coupled receptors (CB1 and CB2) to create live-cell cannabinoid reporter assays. The assays were used to compare activity of synthetic cannabinoids and their metabolites and also to screen urine for CB receptor activity using the Nano-Glo® Live Cell Assay System. (4965)

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Sci. Rep. 6, 29130. Determination of GLUT1 oligomerization parameters using bioluminescent Förster resonance energy transfer. 2016

Looyenga, B., VanOpstall, C., Lee. Z., Bell, J., Lodge, E., Wrobel, K., Arnoys, E. and Louters, L.

Notes: Oligomerization of the glucose transporter (GLUT1) within this plasma membrane was assessed using the NanoBRET™ system. When expressed at high levels, GLUT1 has been shown to form tetrameric complexes with higher transport efficiency. Theoretical NanoBRET™ efficiency was determined for a variety of fluorescent protein acceptors and experiments were performed with mCherry as the NanoBRET™ acceptor. GLUT1 was shown to form a range of higher order complexes in live cells. Flow cytometry and immunoblotting were used in parallel to estimate GLUT1 density in cells. (5054)

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PLos ONE 11, e0161930. Development of a Quantitative BRET Affinity Assay for Nucleic Acid-Protein Interactions 2016

Vickers, T.A. and Crooke, S.T.

Notes: Proteins fused to NanoLuc® luciferase were used as the energy donor and AlexaFluor conjugated nucleic acids were used as the energy acceptor in BRET assays that measured protein:nucleic acid interactions. The assay was demonstrated with immunopurified proteins, cell homogenates, and in whole cells. NanoLuc® fusion protein construction, expression, and purification were performed using the vectors pFN31K Nluc CMV-neo for amino-terminal clones and pFC32K Nluc CMV-neo for carboxy-terminal clones. BRET assays were performed in white 96-well plates. Alexa-linked anti-sense oligonucleotides at the indicated concentrations were incubated at room temperature for 15 min in 1X binding buffer with 106 RLU/well of immunoprecipitated NLuc fusion protein or whole cell lysate. Following the incubation, NanoGlo® substrate was added at 0.1 μl/well. Readings were performed using a GloMax® Discover System. (4757)

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Mol. Cancer Ther. 15, [ePub ahead of print]. Development of a RSK Inhibitor as a Novel Therapy for Triple Negative Breast Cancer 2016

Ludwik, K.A., Campbell, J.P., Li, M., Li, Y., Sandusky, Z.M., Pasic, L., Sowder, M.E., Brenin, D.R., Pietenpol, J.A., O'Doherty, G.A. and Lannigan, D.A.

Notes: 2D and 3D proliferation assays were performed using the CellTiter-Glo® Reagent with a GloMax® Discover System. (4745)

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eLife 5, (epub ahead of print) e10557. Discovery and validation of sub-threshold genome-wide association study loci using epigenomic signatures. 2016

Wang, X., Tucker, N.R., Rizki, G., Mills, R., Krijger, P.H.L., de Wit, E., Subramanian, V., Bartell, E., Nguyen, X.-X., Ye, J., Leyton-Mange, J., Dolmatova, E.V., van der Harst, P., de Laat, W., Elinor, P.T., Newton-Cheh, C., Milan, D.J., Kellis, M. and Boyer, L.A.

Notes: Putative enhancer elements were cloned into the pGL4.23 [luc2/minP] and transfected into human iCMs (Cellular Dynamics) along with a Renilla luciferase control. The iCMs were plated at 2 × 103 per well of a 96-well plate and cultured until all cells were electrically connected and beat simultaneously (~7 days post plating). Beating cells were transfected with the two reporters using the ViaFect™ Transfection Reagent at a 2:1 Viafect:DNA ratio (further detail provided in the paper). The reporter activities were measured 24 hours post-transfection with the Dual-Luciferase® Reporter Assay System. Transfections using a GFP-expressing vector provided a visual estimation of the transfection efficiency and 65-70% of the iCMs were GFP-positive 24 hrs post-transfection. (4671)

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Genes Cells 21(1), 25–40. Distal regulatory element of the STAT1 gene potentially mediates positive feedback control of STAT1 expression. 2016

Yuasa, K. and Hijikata, T.

Notes: The function of the regulatory element, 5.5URR, upstream of signal transducer and activator of transcription 1 (STAT1) is investigated. The Dual-Luciferase® Reporter Assay showed a significant increase in transcription in the presence of the 5.5URR upon interferon treatment. The HaloCHIP™ System showed a physical interaction between the 5.5URR element and the STAT1 promoter, which was stimulated by interferon treatment. Together, the 5.5URR element may serve in maintaining interferon signaling in relation to STAT1. (5097)

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Biochim. Biophys. Acta 1863, 40–9. DNA damage-induced apoptosis suppressor (DDIAS), a novel target of NFATc1, is associated with cisplatin resistance in lung cancer. 2016

Im, J.-Y., Lee, K.-W., Won, K.-J., Kim, B.-K, Ban, H.S., Yoon, S.-H., Lee, Y.-J., Kim, Y.-J., Song, K.-B. and Won, M.

Notes: The functional elements of the DDIAS promoter were deciphered with the Dual-Luciferase® Reporter Assay System utilizing constructs in pGL2 Basic and pRL-TK Control Vectors. NCI-H1703 cells were treated with siRNAs to either DDIAS, NFATc1 or a scrambled sequence. The cells with the scrambled sequence were unchanged, but the other two induced cytotoxicity and caspase-3/7 activation. Cytotoxicity was assessed with the CellTox™ Green Cytotoxicity Assay, and caspase-3/7 was assessed with a kinetic reagent. The two assays were visualized and quantified with an IncuCyte ZOOM System. (4707)

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Mol. Cancer Ther. 15, 2119–29. Dual and specific inhibition of NAMPT and PAK4 by KPT-9274 decreases kidney cancer growth. 2016

Abu Aboud, O., Chen, C.-H., Senapedis, W., Blaglu, E., Argueta, C. and Weiss, R.H.

Notes: The effect of KPT-9274 on RPTEC, Caki-1 and 786-0 cell NAD+/NADH levels was assessed with the NAD/NADH-Glo™ Assay. The treatment was performed on 3,000 cells per well for 48 hours prior to assay. (4840)

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