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PLoS Pathogens 14, e1007057. Plasmodium parasite exploits host aquaporin-3 during liver stage malaria infection. 2018

Posfai, D., Sylvester, K., Reddy, A., Ganley, J.G., Wirth, J., Cullen, Q.E., Dave, T., Kato, N., Dave, S.S., and Derbyshire, E.R.

Notes: Researchers studied the liver stage of the Plasmodium parasite (which causes malaria), including RNA sequencing, parasite load and cell viability. (5035)

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Stem Cells 36, 55–64. Pluripotent stem cell‐derived hematopoietic progenitors are unable to downregulate key epithelial‐mesenchymal transition‐associated miRNAs 2018

Meader, E., Barta, T., Melguizo-Sanchis, D., Tilgner, K., Montaner, D., El-Harouni, A.A., Armstrong, L. and Lako, M.

Notes: Reliaprep RNA cell miniprep system was used to extract RNA from Hematopoietic stem cells derived from pluripotent stem cells. Then qRT-PCR analysis of miRNA and reverse transcription was performed using non-Promega products and 1 μg of RNA was used in a 20 μl GoScript reverse transcription reaction for the analysis of gene expression by qRT-PCR. (4975)

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Stem Cells 36, 337–348. Differences in the Activity of Endogenous Bone Morphogenetic Protein Signaling Impact on the Ability of Induced Pluripotent Stem Cells to Differentiate to Corneal Epithelial‐Like Cells 2017

Kamarudin, T.A., Bojic, S., Collin, J., Yu, M., Alharthi, S., Buck, H., Shortt, A., Armstrong, L., Figueiredo, F.C., Lako, M.

Notes: RNA was extracted from the cells collected from differentiating hESC and hiPSC at days 0, 9, and 20 using the ReliaPrep RNA Cell Miniprep System. The RNA quality was evaluated using the Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, MA). One microgram of extracted RNA was converted into cDNA using reverse transcription (GoScript Transcription System). Quantitative real‐time polymerase chain reaction (qRT‐PCR) was carried out using the QuantStudio 7 Flex Real‐Time PCR System (Thermo Fisher Scientific, MA) and GoTaq qPCR Master Mix. pGL3 BRE Luciferase was used for plasmid lipofection to transfect the cells in each well of a 24-well plate. Cells that were transfected with empty vector (pGL3‐Basic) or BMP reporter (pGL3 BRE Luciferase) were cotransfected with empty Renilla vector (pRL‐Null). Luciferase activities were evaluated with a Dual‐Luciferase Assay System. (4982)

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EMBO Mol Med 9, 482–497. Gene expression profiling of patient‐derived pancreatic cancer xenografts predicts sensitivity to the BET bromodomain inhibitor JQ1: implications for individualized medicine efforts. 2017

Bian, B., Bigonnet, M., Gayet, O. and 25 others.

Notes: Primary pancreatic cancer cell lines were grown a microsphere by culturing in round bottom plates in the presence of methylcellulose. Spheroids were assessed for sensitivity to JQ1 using the CellTiter-Glo® 3D Cell Viability Assay. Cell lines expressing high levels of c-myc were sensitive to JQ1. RNA was isolated from cell lines and targeted expression analysis was performed by reverse transcription with the GoScript™ Reverse Transcription System followed by dye-based qPCR with the GoTaq® qPCR Master Mix on the AriaMx real-time PCR instrument (Agilent). (1825)

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FASEB J. 30(2), 24-34. Cold-inducible RBM3 inhibits PERK phosphorylation through cooperation with NF90 to protect cells from endoplasmic reticulum stress. 2016

Zhu, X., Zelmer, A., Kapfhammer, J.P., and Wellmann, S.

Notes: HEK 293 cells were treated with thapsigargin and expression of target mRNAs was examined. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reverse transcribed with the GoScript™ Reverse Transcription System followed by quantitation with the dye-based GoTaq® qPCR System. (4606)

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Microbiol Immunol. 60, 17-25. Interaction of the hepatitis B virus X protein with the lysine methyltransferase SET and MYND domain-containing 3 induces activator protein 1 activation. 2016

Hyashi, M., Deng, L., Chen, M., Gan, X., Shinozaki, K., Shoji, I., and Hotta, H.

Notes: The effect of SMYD3 and ERK2 on hepatitis B RNA expression was examine in transfected Huh-7.5 human hepatoma cells. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System followed by dye-based qPCR analysis. The effect of SMYD3 and ERK2 on AP1-dependent and NF-κB-dependent gene expression in the cells was examined with the pGL4.44 [luc2P/AP1 RE/Hygro] and pGL4.32 [luc2P/NF-κB-RE/Hygro] Reporter vectors using the Dual-Luciferase® Reporter Assay System with a pRL-TK Renilla luciferase control. Reporter assays were read with a GloMax® Instrument. (4603)

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J. Virol. Methods 236, 221-230. Towards next-generation sequencing analytics for foodborne RNA viruses: Examining the effect of RNA input quantity and viral RNA purity.  2016

Yang Z., Leonard S., Mammel M., Elkins C., Kulka M.

Notes: In this study, researchers treated selected viral RNA samples with 1 unit of DNase I to deplete the presence of any genomic DNA in the extracts. The viral RNA genomic regions of interest were reverse transcribed into cDNA using oligo(dT) primer and AMV reverse transcriptase. (4887)

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BMC Cancer 15, 634. Chemotherapy induces Notch1-dependent MRP1 up-regulation, inhibition of which sensitizes breast cancer cells to chemotherapy. 2015

Kim, B., Stephen, S.L., Hanby, A.M., Horgan, K., Perry, S.L., Richardson, J., Roundhill, E.A., Valleley, E.M.A., Verghese, E.T., Williams, B.J., Thorne, J.L. and Hughes, T.A.

Notes: T47D and HB2 breast cancer cell lines were treated with doxorubicin and expressed in three target genes. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System, reverse transcribed with the GoScript™ Reverse Transcription System and analyzed with the dye-based GoTaq® qPCR System. (4612)

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Cancer Lett. 356, 994-1006. Lobatin B inhibits NPM/ALK and NF-κB attenuating anaplastic-large-cell-lymphomagenesis and lymphendothelial tumour intravasation. 2015

Kiss, I., et al.

Notes: SR-786 NPM/ALK-positive human anaplastic large cell lymphoma cell line was treated with lobatin B, a plant-derived natural compound. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reversed transcribed into cDNA with the GoScript™ Reverse Transcription System. Levels of NPM/ALK and NF-κB expression was measured with the dye-based GoTaq® qPCR System. SR-786 cells were also analyzed for functioning metabolism with the CellTiter-Blue® Cell Viability Assay and apoptosis with the Apo-ONE™ Homogeneous Caspase-3/7 Assay. (4604)

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Mediators Inflammation , Article ID: 737310. Role of calprotectin as a modulator of the IL27-mediated proinflammatory effect on endothelial cells. 2015

Dorosz, S.A., Ginolhac, A., Kähne, T., Naumann, M., Sauter, T., Salsmann, A., and Bueb, J.-L.

Notes: Human Umbilical Vein Endothelial cells (HUVECs) were treated with IL-27, calprotectin, TNFα or combinations. Cell number per well was controlled by lysing the cells and measuring the LDH release with the CytoTox 96® Assay. Changes in gene expression were monitored by probe-based qPCR with cDNA made with the GoScript™ Reverse Transcription System using total RNA isolated using the ReliaPrep™ RNA Cell Miniprep System. Changes in protein levels upon treatment were evaluated by mass spec using Trypsin Gold for peptide analysis. (4598)

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Antiviral Res. 124, 69–76. The non-pathogenic Henipavirus Cedar paramyxovirus phophoprotein has a compromised ability to target STAT1 and STAT2. 2015

Lieu, K.G., Marsh, G.A., Wang, L.-F. and Netter, H.J.

Notes: HEK 293 cells were transfected with various viral protein expression vectors and expression levels measured in the presence or absence of interferon-α using RT-qPCR. Total RNA was isolated from the cells using the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System prior to dye-based qPCR. (4613)

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PLos ONE 9, e106190. Anoctamin-1 in the juvenile rat urinary bladder. 2014

Bijos, D.A., Drake, M.J., and Vahabi, B.

Notes: Total RNA was extracted from mucosa and denuded-detrusor layers of rat bladders with the ReliaPrep™ RNA Tissue Miniprep System and converted to cDNA using GoScript™ Reverse Transcription System prior to PCR amplification. (4608)

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Mol. Cell. Biol. 34, 3486–99. Cellular migration and invasion uncoupled: Increased migration is not an inexorable consequence of epithelial-to-mesenchymal transition. 2014

Schaeffer, D., Somarelli, J.A., Hanna, G. and Palmer, G.M.

Notes: Rat renal DT and A3 cells were examined for differences in EGFR expression by RT-qPCR. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reverse transcribed with the GoScript™ Reverse Transcription System followed by dye-based qPCR analysis. (4617)

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PLos ONE 9, e98877. Induction of cell-mediated immune responses in mice by DNA vaccines that express hepatitis C virus NS3 mutants lacking serine protease and NTPase/RNA helicase activities. 2014

Ratnoglik, S.L., Jiang, D.-P., Aoki, C., Sudarmono, P., Shoji, I., Deng, L. and Hotta, H.

Notes: Primary splenocytes were isolated from mice immunized with an NS3 expression vector and examined for IFN-γ expression by RT-qPCR. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System. The affect of wildtype and mutant NS3 on interferon-β promoter activity with an IFN-β promoter firefly luciferase vector and pRL-TK vector control was measured with the Dual-Luciferase® Reporter Assay System. LDH release was measured with the CytoTox 96® Cytotoxicity Assay in a cytotoxic T-lymphocyte assay using cultured splenocytes from the immunized mice. (4609)

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Fitoterapia. 99, 276–83. Inhibition of hepatitus C virus replication by chalepin and pseudane IX isolated from Ruta angustifolia leaves. 2014

Wahyuni, T.S., Widyawaruyanti, A., Lusida, M.I., Fuad, A., Soetjipto, Fuchino, H., Kawahara, N., Hayashi, Y., Aoki, C. and Hotta, H.

Notes: Huh7.5 cells were infected with HCV and treated with the plant-derived chemicals. The expression of viral messages was analyzed by RT-qPCR. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reverse transcribed with the GoScript™ Reverse Transcription System before dye-based qPCR analysis. (4616)

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Acta Biomater. 10, 3177-87. Three-dimensional hydrogel scaffolds facilitate in vitro self-renewal of human skin-derived precursors. 2014

Wang, X., Liu, S., Zhao, Q., Li, N., Zhang, H., Zhang, X., Lei, X., Zhao, H., Deng, Z., Qiao, J., Cao, Y., Ning, L., Liu, S., and Duan, E.

Notes: Human Skin Progenitor (HSK) cells were cultured on hydrogels and either left alone or differentiated into various skin cell types. Gene expression changes were monitored by isolating total RNA from collagenase and hyaluronidase treated hydrogels with the ReliaPrep™ RNA Cell Miniprep System followed by cDNA synthesis with the GoScript™ Reverse Transcription System and qPCR with the dye-based GoTaq® qPCR System. (4602)

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Nucl. Acids Res. 41, 8515–25. Pin1 promotes GR transactivation by enhancing recruitment to target genes. 2013

Poolman, T.M., Farrow, S.N., Matthews, L., Loudon, A.S. and Ray, D.W.

Notes: RNA was extracted from cultured A549 cells and used in RT-qPCR to analyze the effect of transfected siRNAs. (4444)

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PLos ONE 7, e48183. Enhanced expression of vacuolar H+-ATPase subunit E in the roots is associated with the adaptation of Broussonetia papyrifera to salt stress. 2012

Zhang, M., Fang, Y., Liang, Z. and Huang, L.

Notes: The authors examined cellular adaptation to increased salinity in Broussonetia papyrifera by measuring protein and mRNA levels of vacuolar H+-ATPase (V-H+-ATPase) subunits and the activities of V-H+-ATPase and vacuolar H+-pyrophosphatase. Relative expression levels of V-H+-ATPase subunits A, B, E and c in salt-stressed and control plants were determined by RT-PCR using actin as a normalization control gene. cDNA was synthesized using the GoScript™ Reverse Transcription System as described by the manufacturer’s protocol, then amplified by PCR for 25 cycles. The amplification products were analyzed and quantified by agarose gel electrophoresis and ethidium bromide staining. (4258)

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J. Biol. Chem. 286, 42690-42703. Alternative Splicing Produces Nanog Protein Variants with Different Capacities for Self-renewal and Pluripotency in Embryonic Stem Cells. 2011

Das, S., Jena, S., and Levasseur, D.N.

Notes: The transcription factor Nanog is required for the maintenance of embryonic stem (ES) cell pluripotency. These authors showed that the Nanog N-terminal domain is regulated by post-transcriptional modification, and that alternative splicing generates Nanog variants with different capacities for maintaining an undifferentiated cell state. As part of their study, the authors used GoScript® Reverse Transcriptase to generate cDNA from RNA extracted from cell lines expressing different Nanog variants. The cDNA was used in RT-qPCR to quantify relative expression levels. (4184)

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