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J. Biol. Chem. 273, 25472-25479. Cloning and characterization of the human PAX2 promoter 1998

Stayner, C.K., Cunliffe, H.E., Ward, T.A., Eccles, M.R.

Notes: Promoter functions were tested in NIH 3T3, COS-7, 293 and MDCK cells. Experimental constructs were prepared in the pGL2 Basic Vector and cotransfected with the pRL-TK Vector at a 40:1 ratio. Luciferase activities were monitored with the Dual-Luciferase® Reporter Assay System. (0332)

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J. Biol. Chem. 273, 7643-7649. Cloning of the multipartite promoter of the sodium-calcium exchanger gene NCX1 and characterization of its activity in vascular smooth muscle cells. 1998

Scheller, T., Kraev, A., Skinner, S., Carafoli, E.

Notes: Promoter activities were examined in primary rat vascular smooth muscle cells. The firefly luciferase vectors were co-transfected with the pRL-TK Vector at a 20:1 ratio. Transfections were performed with an unspecified member of the Tfx™ Reagent trio. (0451)

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J. Immunol. 160, 273. Common and distinct signaling pathways mediate the induction of TNF-alpha and IL-5 in IgE plus antigen-stimulated mast cells. 1998

Csonga, R., Prieschl, E.E., Jaksche, D., Novotny, V., Baumruker, T.

Notes: Luciferase reporter plasmids were cotransfected with pRL-TK Vector at a 5:1 ratio in CPII mouse mast cells. In studies with the reporter vector, ras expression vector and pRL-TK Vector, the ratio of each was 3:5:1 with the amount of pRL-TK Vector constant in all transfections at 2µg per reaction. All transfections were accomplished by electroporation and both luciferase activities were determined with the Dual-Luciferase® Reporter Assay System. (1288)

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Immunity 9, 871-880. Direct suppression of Stat1 function during adenoviral infection. 1998

Look, D.C., Roswit, W.T., Frick, A.G., Gris Alevy, Y., Dickhaus, D.M., Walter, M.J., Holtzman, M.J.

Notes: Used Dual-Luciferase® Reporter Assay System with a 400:1 transfection ratio of the firefly luciferase plasmid to pRL-SV40 Vector. Also used the pCI-neo Mammalian Expression Vector to make several E1A expression plasmids. (0743)

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J. Biol. Chem. 273, 26969-26976. Down-regulation of cytochrome P450 1A1 gene promoter by oxidative stress: Critical contribution of nuclear factor 1. 1998

Morel, Y., Barouki, R.

Notes: Promoter activities were studied in HepG2 cells with the Dual-Luciferase® Reporter Assay System. The Renilla control vector was constructed from the pRL-SV Vector by replacing the SV40 promoter with the α-globin gene proximal promoter (–79-+1). The Firefly luciferase construct was prepared in the pGL3 Basic Vector. The pGL3-based Vector and pRL-based Vector were transfected at a 3:1 ratio. A mutant nuclear factor 1 site was introduced into the 1A1 promoter with the GeneEditor™ in vitro Site-Directed Mutagenesis System by converting GCCA to CGCA. (0659)

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Alcohol Clin. Exp. Res. 22, 849-853. Ethanol transcriptionally upregulates t-PA and u-PA gene expression in cultered human endothelial cells. 1998

Grenett, H.E., Aikens, M.L., Torres, J.A., Denissle, S., Tabengwa, E.M., Davis, G.C., Booyse, F.M.

Notes: Studies were performed in low passage HUVEC cells. Two micrograms of pGL2 Basic-derived construct was cotransfected with 0.15µg of the pRL-TK Vector (13:1 ratio). The luciferase activity was determined with the Dual-Luciferase® Reporter Assay System. (1120)

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Mol. Cell 1, 905-911. Functional analysis of the human TAFII250 N-terminal kinase domain 1998

O'Brien, T., Tijan, R.

Notes: Studies were performed in ts13 cells that have a temperature-sensitive TAFII250. Either cyclin A or cdc2 luciferase vectors were cotransfected with wildtype or mutant TAFII250 proteins as well as pRL-CMV Vector (luciferase vector: Renilla vector; 20:1). Luciferase activity was measured with the Dual-Luciferase® Reporter Assay System (0586)

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J. Immunol. 161, 3822. GATA-3-dependent enhancer activity in IL-4 gene regulation. 1998

Ranganath, S., Ouyang, W., Bhattarcharya, D., Sha, W.C., Grupe, A., Peltz, G., Murphy, K.M.

Notes: Three plasmids were transfected into M12 cells or Jurkat cells: Cytokine promoter-driven firefly luciferase reporter vector; GATA-3 expression vector and pRL-CMV Vector. The vectors were electroporated at a 40:40:1 ratio. Luciferase activity was determined with the Dual-Luciferase® Reporter Assay System. (0497)

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J. Biol. Chem. 273, 25466-25471. Heat shock factor 1 mediates hemin-induced hsp70 gene transcription in K562 erythroleukemia cells 1998

Yoshima, T., Yura, T., Yanagi, H.

Notes: Reporter studies were performed in K562 cells. The Dual-Luciferase® Reporter Assay System was used to measure the firefly luciferase from a GAL4 binding site-containing luciferase vector and cotransformed pRL-SV40 Vector. No ratios were reported. Other vectors were transfected as well and produced proteins that activated that bound the GAL4 domain and activated transcription of the firefly luciferase in a two-hybrid-type assay. Another vector was constructed with Heat Shock Elements instead of GAL4 domains in a pGL3 Promoter Vector. (0116)

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J. Biol. Chem. 273, 33741. Identification of the cis-acting endoplasmic reticulum stress response element responsible for transcriptional induction of mammalian glucose-regulated proteins: Involvement of basic leucine zipper transcription factors. 1998

Yoshida, H., Haze, K., Yanagi, H., Yura, T., Mori, K.

Notes: Promoter studies were performed in HeLa cells. The promoter constructs were assembled in the pGL3 Basic Vector (10µg) and cotransformed with the pRL-SV40 Vector (0.1µg) at a 10:1 ratio using the calcium phosphate method. Reporter activity was measured with the Dual-Luciferase® Reporter Assay System. The TNT® T7 Quick Coupled Transcription/Translation System was used to produce a positive control protein for Western blot analysis of transfected HeLa cell extracts. (0115)

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Am. J. Physiol. 274, C681-C687. In vivo analysis of the myosin heavy chain IIB promoter region. 1998

Swoap, S. J.

Notes: The authors cloned a 2.6kb promoter-enhancer region of the MHC IIB gene into the pGL3-Basic Vector. They injected this construct along with the pRL-CMV Vector into mouse muscle cells. Tissue extracts were prepared with a buffered Tris solution containing a protease inhibitor cocktail and assayed for reporter activity using the Dual-Luciferase® Reporter Assay System. (0287)

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J. Biol. Chem. 273, 22120-22127. Induction of cyclooxygenase-2 by activated ha-ras oncogene in Rat-1 fibroblasts and the role of mitogen-activated protein kinase pathway. 1998

Sheng, H., Williams, C.S., Shao, J., Liang, P., DuBois, R.N., Beauchamp, R.D.

Notes: Luciferase studies were performed in a Rat-1 fibroblast cell line with an inducible Ha-ras gene. Experimental pGL3 Basic vectors were cotransfected with pRL-TK at a 2:1 ratio. (0389)

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J. Biol. Chem. 273, 22877-22883. Insulin-like growth factor-I augments erythropoietin-enduced proliferation through enhanced tyrosine phosphorylation of STAT5. 1998

Okajima, Y., Matsumura, I., Nishiura, T., Hashimoto, K., Yoshida, H., Ishikawa, J., Wakao, H., Yoshimura, A., Kanakura, Y., Tomijama, Y., Matsuzawa, Y.

Notes: Studies were performed in F-36P, a human IL-3 dependent erythroleukemia cell line. Cells were electroporated with either an AP-1 specific luciferase reporter vector or a STAT5-specific luciferase reporter vector. To normalize for electroporation efficiency, an equal amount of the pRL-CMV Vector was co-transfected with the luciferase reporters. Luciferase activities were monitored with the Dual-Luciferase® Reporter Assay System. (0599)

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J. Biol. Chem. 273, 13552-13562. Isoforms of hepatocyte nuclear factor-6 differ in DNA-binding properties, contain a bifunctional homeodomain and define the new ONECUT class of homeodomain proteins. 1998

Lannoy, V.J., Bürglin, T.R., Rousseau, G.G., Lemaigre, F.P.

Notes: Studies were performed in HepG2 cells. Cells were transfected with a hepatocyte nuclear factor-6 (HNF-6)–responsive promoter firefly luciferase construct (3µg), various forms of the HNF-6 protein in an expression vector (400ng) and a Renilla luciferase control vector (pRL-null Vector) driven by the liver pfk-2 promoter (not responsive to HNF-6; 500ng). The ability of the various HNF-6 isoforms to drive luciferase expression was normalized to Renilla luciferase activity, and the luciferase activity was measured with the Dual-Luciferase® Reporter Assay System. The various HNF-6 constructs were also expressed in vitro with the TNT® Coupled Wheat Germ Extract System and used for gel shift assays. (0839)

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J. Biol. Chem. 273, 8278-8286. Lymphoid-specific expression of the Id3 gene in hematopoietic cells. Selective antagonism of E2A basic helix-loop-helix protein associated with Id3-induced differentiation of erythroleukemia cells. 1998

Deed, R. W. , Jasiok, M. , Norton, J. D.

Notes: Promega's Taq DNA polymerase was used in construction of various plasmids. The authors used a mammalian two-hybrid system with a firefly luciferase reporter and pRL-CMV Vector as a transfection control. Luciferase activity was measured using the Dual-Luciferase® Reporter Assay System. (1266)

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Circ. Res. 83, 508-515. Lysophosphatidylcholine enhances cytokine-induced interferon gamma expression in human T lymphocytes. 1998

Nishi, E., Kume, N., Ueno, Y., Ochi, H., Moriwaki, H., Kita, T.

Notes: Promoter studies were performed in human peripheral T lymphocytes. Experimental constructs in the pGL2-Basic Vector and cotransfected with the pRL-TK Vector at a 20:1 ratio. The luciferases were detected with the Dual-Luciferase® Reporter Assay System. (0616)

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J. Biol. Chem. 273, 19130-19139. Phorbol ester-induced transcription of a fibroblast growth factor-binding protein is modulated by a complex interplay of positive and negative regulatory promoter elements. 1998

Harris, V.K., Liaudet-Coopman, E.D.E., Boyle, B.J., Wellstein, A., Riegel, A.T.

Notes: Reporter studies were performed in the ME-180 squamous cell carcinoma cell line. One microgram of the firefly luciferase construct was contransfected with 1ng of pRL-CMV Vector (1,000:1 ratio of firefly reporter to Renilla control), and luciferase activities were measured with the Dual-Luciferase® Reporter Assay System. The Kanamycin Control RNA was used as a control template for primer extension analysis. (1065)

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Nature 391, 82-86. PPAR-γ agonists inhibit production of monocyte inflammatory cytokines. 1998

Jiang, C., Ting, A.T., Seed, B.

Notes: The authors used the Dual-Luciferase® Reporter System with U937 cells and the pRL-TK Vector. (0977)

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Proc. Natl. Acad. Sci. USA 95, 8170-8174. Racial variability in the UDP-glucuronosyltransferase 1 (UGT1A1) promoter: A balanced polymorphism for regulation of bilirubin metabolism? 1998

Beutler, E., Gelbart, T. and Demina, A.

Notes: Studies were performed in HepG2 and HuH7 cells (both human cells of hepatic origin). One microgram of the experimental constructs in the pGL3 Basic Vector was cotransfected with 50ng of the pRL-SV Vector (20:1 ratio). Luciferase activity was measured using the Dual-Luciferase® Reporter Assay System. (1443)

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J. Biol. Chem. 273, 27474-27483. Sp1 and Sp3 regulate transcriptional activity of the facilitative glucose transporter isoform-3 gene in mammalian neuroblasts and trophoblasts. 1998

Rajakumar, R.A., Thamotharan, S., Menon, R.K., Devaskar, S.U.

Notes: AMV Reverse Transcriptase was used for primer extension analysis from poly(A)+ RNA. The Riboprobe® in vitro Transcription System was used to generate [32P]-antisense RNA probes for RNase protection assays. Promoter studies were performed in N2A murine neuroblasotoma cells. Experimental constructs were assembled in the pGL2-Basic Vector and cotransfected with the pRL-TK Vector at a 10:1 ratio and luciferase activities determined with the Dual-Luciferase® Reporter Assay System. Some promoter studies were performed in Drosophila Schneider cells with luciferase promoter constructs and an RSV-driven beta-galactosidase vector. Beta-Galactosidase activity was determined with the Beta-Galactosidase Enzyme Assay System. (0531)

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J. Biol. Chem. 273, 26923-26930. The early growth response protein (EGR-1) regulates interleuking-2 transcription by synergistic interaction with the nuclear factor of activated T cells. 1998

Decker, E.L., Skerka, C., Zipfel, P.F.

Notes: The 293 cell line was cotransfected with a luciferase reporter containing the IL-2 promoter, an expression vector for either EGR-1, NFATc or both and finally the pRL-TK Vector. Reporter to control ratio was 4:1 and the luciferase activity was determined with the Dual-Luciferase® Reporter Assay System. (1265)

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J. Biol. Chem. 273, 34775. The roles of nuclear factor of activated T cells and Ying-Yang 1 in activation-induced expression of the interferon-gamma promoter in T cells. 1998

Sweetser, M.T., Hoey, T, Sun, Y.L., Weaver, W.M., Price, G.A., Wilson, C.B.

Notes: Reporter assays were performed in primary murine splenocytes. The interferon-gamma promoter constructs were assembled in the pGL3-Basic Vector and a Renilla luciferase control vector was constructed with the pRL-null Vector and a β-actin promoter. The plasmids were electroporated at a 10:1 ratio. Luciferase activities were followed with the Dual-Luciferase® Reporter Assay System. (0285)

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J. Lipid Res. 39, 1520-1524.. The –514 polymorphism in the hepatic lipase gene (LIPC) does not influence androgen-mediated stimulation of hepatic lipase activity. 1998

Vega, G.L., Gao, J., Bersot, T.P., Mahley, R.W., Verstraete, R., Grundy, S.M., White, A., Cohen, J.C.

Notes: Studies were performed in HepG2 cells. Experimental promoter contructs were assembled in the pGL3-Basic Vector and transfection efficiency was monitored by cotransfecting the pRL-CMV Vector. Luciferase activities were assayed with the Dual-Luciferase® Reporter Assay System. The ratio of experimental:control vector were not reported. (0219)

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Biochem. J. 332, 773-780. Transcript heterogeneity of the human reduced folate carrier results from the use of multiple promoters and variable splicing of alternative upstream exons. 1998

Zhang, L. , Wong, S. C. , Matherly, L. H.

Notes: The authors used the PolyATtract® mRNA Isolation System to isolate mRNA from total RNA. They used this mRNA-rich fraction for primer extension analysis using Promega's AMV Reverse Transcriptase. The Wizard® Plus Midiprep DNA Purification System was used for various plasmid isolations and the Erase-a-Base®  System was used to generate deletion series. The Dual-Luciferase® Reporter Assay System was used to study promoters cloned into the pGL3-Basic Vector. The pRL-SV40 was used as a transfection control plasmid. (0095)

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J. Biol. Chem. 273, 21137-21144. Transcriptional activation of the p21WAF1, CIP1, SDI1 gene by interleukin-6 type cytokines: A prerequisite for their pro-differentiating and anti-apoptotic effects on human osteoblastic cells. 1998

Bellido, T., O'Brien, C.A., Roberson, P.K. and Manolagas, S.C.

Notes: Studies were performed in MG63 human osteosarcoma cells. Firefly luciferase activity was normalized to Renilla luciferase activity using the Dual-Luciferase® Reporter Assay System. The cells were transfected with the pGL2-Basic Vector containing the experimental promoters, pRL-CMV Vector, and a Stat-3 expression vector. The ratio of pGL2 Vector to pRL-CMV Vector was 1000:1. The ratio of pGL2 Vector to Stat3 Expression Vector was 6:1. The chemical MTS was used in a LDH-coupled assay to determine cell viability of osteosarcoma cells transfected with antisense oligonucleotides. (1436)

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