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J. Biol. Chem. 278 (10), 8018-8027. Nuclear factor-κB and mitogen-activated protein kinases mediate nitric oxide-enhanced transcriptional expression of interferon-kβ. 2003

Jacobs, A.T. and Ignarro, L.J.

Notes: Promega T4 Polynucleotide Kinase was used to end-label an oligo representing a NF-kB binding sequence element with [γ-32P] ATP (7,000 Ci/mmol).  Specific primers to IFN-β and IκB-α messenger RNA were used in RT-PCR to generate products for cloning into the pGEM®-T Easy Vector. The resulting plasmids were linearized with Nco I and Spe I, respectively, and used as templates for in vitro transcription using the Riboprobe® System to generate probes for use in an RNase protection assay.  The antisense probes were labeled with [α-32P]CTP (800 Ci/mmol) in the Riboprobe® reactions.  RNase ONE™ Ribonuclease was also used in this study.  (3197)

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J. Cell Sci. 116, 4467-4478. Organization and translation of mRNA in sympathetic axons. 2003

Lee, S.K. and Hollenbeck, P.J.

Notes: In situ hybridization was performed on 4% formalin-fixed chick sympathetic neurons.  As a control for specificity in the in situ hybridization experiments, samples were treated with RNase ONE Ribonuclease.  Samples were incubated with RNase ONE Ribonuclease for 30-120 minutes at 37°C before the probe hybridization step.  (3196)

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EMBO J. 19(1), 124-33. Replicase activity of purified recombinant protein P2 of double-stranded RNA bacteriophage phi6. 2000

Makeyev, E.V., Bamford, D.H.

Notes: The authors performed an RNAse protection using RNAse ONE™ Ribonuclease. The reaction was performed at 28°C for one hour and stopped by addition of 0.1% SDS and 10µg E. coli tRNA. The template used was RNA synthesized by the Phi6 phage P2 polymerase. The protection was done as part of a strand separation analysis to determine if the newly synthesized RNA is double stranded, composed of a newly synthesized RNA and the original template RNA. (2209)

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Mol. Cell. Biol. 19, 274-283. Transcriptional and posttranscriptional silencing of rodent α1(I) collagen by a homologous transcriptionally self-silenced transgene. 1999

Bahramian, M.B. and Zarbl, H.

Notes: RNase ONE™ Ribonuclease was used for RNase protection assays. The authors note that when they used a mixture of RNase A and T1 in place of RNase ONE™ Ribonuclease they had to be more diligent with their assays because the RNase A/T1 mixture was more likely to digest the protected fragment. RNase ONE™ Ribonuclease demonstrated single-stranded specificity in their experiments. (1492)

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Proc. Natl. Acad. Sci. USA 95, 8767-8772. alpha-Fetoprotein gene sequences mediating Afr2 regulation during liver regeneration. 1998

Jin, D.K., Vacher, J. and Feuerman, M.H.

Notes: These authors used Promega's RNase ONE™ Ribonuclease for RNase Protection Analysis (RPA's). (2031)

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Oncogene 16, 423-428. C-Myc 5' untranslated region contains an internal ribosome entry segment. 1998

Stoneley, M., Paulin, F. E., Le Quesne, J. P., Chappell, S. A., Willis, A. E.

Notes: The authors used the Dual-Luciferase® Reporter Assay System, pGL3 Control Vector, pRL-CMV Vector, pSV-beta-Galactosidase Control Vector, Luciferase Assay System and RNase ONE® Ribonuclease in their studies. The objective of the paper was to demonstrate that the 5' untranslated region (UTR) of c-myc functions as an internal ribosome entry site (IRES). To directly assay the ability of the UTR to function as a IRES, the Renilla luciferase gene was subcloned in front of the UTR. Dual expression of the luciferases was demonstrated in both HeLa cells and HepG2 cells and dependent upon the c-myc UTR. Mutation of the Renilla luciferase sequence to contain a hairpin structure, demonstrate that the Renilla luciferase activity could reduced by 75% with no effect on the firefly luciferase activity. RNase protection assays with a 624 nucleotide probe that crossed the Renilla luciferase cDNA, the c-myc UTR and the firefly luciferase cDNA demonstrated that only one single message is produced by the transfected HeLa cells and the c-myc UTR is not functioning as a promoter. All transfections were normalized to control β-galactosidase activity. (0339)

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J. Biol. Chem. 273, 27420-27429. Characterization of the muring fatty acid transport protein gene and its insulin response sequence. 1998

Hui, T.Y., Frohnert, B.I., Smith, A.J., Schaffer, J.E., Bernlohr, D.A.

Notes: Luciferase reporter constructs were cotransfected into 3T3-L1 preadipocytes with the pRL-CMV Vector at a 25:1 ratio and luciferase activities were determined with the Dual-Luciferase® Reporter Assay System. RT-PCR was accomplished with AMV Reverse Transcriptase and Taq DNA Polymerase. RNase Protection Assay were performed with the aid of the RNase ONE™ Ribonuclease. (0988)

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Biochemistry 36, 5004-5019. Control of complexity constraints on combinatorial screening for preferred oligonucleotide hybridization sites on structured RNA 1997

Bruice, T.W., Lima, W.F.

Notes: In this paper, Promega's RNase ONE® Ribonuclease was used extensively for mapping of RNA interactions. The techniques in the paper are: 1) Determination of binding constants by RNase ONE® Ribonuclease footprinting assay; 2) RNase ONE® Ribonuclease footprinting assay for combinatorial hybridization affinity screening; and 3) RNase ONE™ Ribonuclease footprinting assay, post gel mobility-shift resolution, for combinatorial hybridization affinity screeening. (1369)

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EMBO J. 16, 4082-4091. Identification and characterization of a yeast homolog of U1 snRNP-specific protein C. 1997

Tang, J., Abovich, N., Fleming, M.L., Seraphin, B., Rosbash, M.

Notes: The RNase ONE™ Ribonuclease was used in an RNase I protection assay for mapping U1 snRNP structure. The enzyme was used at a 1:2000 or a 1:4000 dilution. (0301)

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EMBO J. 16, 5006-5018. Multiple circadian-regulated elements contribute to cycling period gene expression in Drosophila 1997

Stanewsky, R., Jamison, C. F., Plautz, J. D., Kay, S. A., Hall, J. C.

Notes: RNase protection assays were performed on 8µg of total Drosophila head RNA toward transgenic constructs using the RNase ONE™ Ribonuclease. Beetle luciferin was used to monitor luciferase activity in vivo. (0331)

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J. Biol. Chem. 272, 5936-5942. Nerve growth factor up-regulates the N-methyl-D-aspartate receptor subunit 1 promoter in PC12 cells. 1997

Bai, G. and Kusiak, J.W.

Notes: Reporter vectors (produced with the pGL2 Basic Vector) were transiently transfected into PC12 cells and promoter activity was measured in response to 2.5S NGF. Promoter constructs were generated by PCR, subcloned into the pGEM®-T Vector and sequenced. The pGEM®-luc Vector was used to generated an antisense luc RNA probe used in RNase protection assays (using RNase ONE™ Ribonuclease) to follow the transcription of luciferase DNA following NGF treatment. The early growth reaction transcription factor and variants were expressed in the TNT® SP6 Coupled Reticulocyte Lysate System and used to perform gel shifts with promoter elements. The SP1 Consensus Oligonucleotide was used to compete transcription factor binding to the promoter elements. (1493)

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