Notes: The RNasin Ribonuclease Inhibitor was used to protect RNA transcripts during primer extension analysis and in vitro transcription of RNA probes. The resultant RNA probes were used for RNase protection assays. (1639)

Notes: The Serine/Threonine Phosphatase Assay System was used to assess protein phosphatase 2A, 2B and 2C activity in nuclear extracts of Human umbilical vein cells (HUVEC) with or without lysophosphatidylcholine treatment. Gel shifts were also performed with nuclear extracts prepared from the HUVEC cells using the SP1 Consensus Oligonucleotide. Luciferase reporter studies were also performed in HUVEC cells using pGL3 Basic-derived vectors and the Luciferase Assay System. (1305)

Notes: A NF-kappaB luciferase reporter vector was constructed with the TATA box and transcription start site of the rabbit beta-globin promoter and three tandem repeats of the NF-kappaB consensus site in the pGL3 Basic Vector. Another vector was constructed with mutated NF-kappaB sites. The reporter vector was transfected into primary human umbilical vein endothelial cells with the Tfx™-50 Reagent. No details of the transfection conditions were provided. Luciferase activities were monitored with the Luciferase Assay System. (0418)

Notes: Luciferase studies were performed in AtT20 cells using the Luciferase Assay System. Experimental constructs were prepared in the pGL3 Basic Vector. (1392)

Notes: The Quantilum™ Recombinant Luciferase was heat denatured in the presence of heat shock protein hsp 18.1 and the renaturation of the protein was assayed in with the Luciferase Assay System. The denaturation was performed in either Rabbit Reticulocyte Lysate or Wheat Germ Extract in the presence or absence of added ATP. (0809)

Notes: Luciferase and CAT assays were conducted in CHO-K1 cells using constructs prepared in the pGL3-Basic and pCAT^®-Basic Vectors, and the activity was normalized to β-galactosidase activity supplied by the pSV-β-Galactosidase Control Vector. The pCAT^® Vectors have been replaced by the next-generation pCAT^®3 Vectors. (1055)

Notes: Luciferase reporter constructs containing either the luc or luc+ gene (from pSP-luc+NF Fusion Vector) were used in in vitro transcription to produce capped mRNA. Luciferase was produced by in vitro translation using either neurospora extract or Promega's Rabbit Reticulocyte Lysate, Nuclease Treated and detected using the Luciferase Assay Reagent. (0208)

Notes: Luciferase studies were performed in CA77 thyroid C cell line UI-0 using constructs prepared in the pGL3-Basic Vector. (0838)

Notes: Luciferase studies were performed in aortic and vena caval smooth muscle cells. Transfections were normalized to β-galactosidase activity. Researchers report that pGL3 Promoter and pGL3 Control Vectors were 55-± 3-fold and 340±41-fold greater activity, respectively, than the pGL3 Basic Vector alone. The CREB consensus oligo was used in gel shifts of aortic and vena cava smooth muscle cell nuclear extracts. (1214)

Notes: Luciferase studies were performed in KRC-7 rat hepatoma cells using constructs prepared in the pGL3 Basic Vector. The luciferase constructs were cotransfected with the pSV-beta-galactosidase Vector. Luciferase activities determined with the Luciferase Assay System were normalized to readings from the Beta-Galactosidase Enzyme Assay System. (1364)

Notes: Reporter studies were performed in HepG2 cells. The experimental constructs were made using the pGL3 Basic Vector and luciferase activities were monitored with the Luciferase Assay System. (0669)

Notes: The Transfectam® Reagent was used to transiently transfect A549 human lung cancer cells. Many details of the transfection are provided. Luciferase activity was normalized to Beta Galactosidase activity. (1240)

Notes: Authors make 3 base changes in a promoter element using the Altered Sites^® II in vitro Mutagenesis System and put the mutated promoter into the pGL3-Basic Vector. Luciferase activity was measured using the Luciferase Assay System. (1047)

Notes: Luciferase studies were performed in 293T cells using constructs prepared in the pGL3 Basic Vector. (1278)

Notes: This paper describes a method for quantitation of luciferase mRNA by in vitro translation using Promega TNT^® Coupled Wheat Germ Extract System. (Rabbit reticulocyte lysate could not be used because of luciferase quenching problems). In this reporter gene assay, COS cells were transfected with a firefly luciferase reporter plasmid driven by promoters/enhancers of varying strengths and total cellular RNA was isolated and translated in vitro using the TNT^® System. To normalize expression, a CMV Renilla luciferase or beta-galactosidase vector was used as a control. To measure luciferase activity either Promega Luciferase Assay System or Dual-Luciferase^® Reporter Assay System was used. (2047)

Notes: Luciferase studies performed in HepG2 cells using constructs prepared in the pGL3 Basic Vector. The luciferase constructs were cotransfected with the pSV-beta-Galactosidase Control Vector to normalize for transfection efficiency. Luciferase activities were measured with the Luciferase Assay System. (0629)

Notes: The pCI-Neo Mammalian Expression Vector was used in conjunction with a luciferase reporter vector to determine the important regions on the carboxy-terminal portion of the 386-amino acid LMP-1 protein. Both full-length and truncation mutants were expressed in the BJAB human EBV-negative lymphoblastoid B cell line. Luciferase activity was measured using the Luciferase Assay System. (1409)

Notes: Calf Intestinal Alkaline Phosphatase was used for in vitro dephosphorylation of the RAP46 protein, and the dephosphorylated protein was assayed for interaction with various transcription factors. The TNT^® Coupled Reticulocyte Lysate System was used to produce the transcription factors. The Rabbit Reticulocyte Lysate was used to aid protein refolding of heat denatured luciferase and the extent of refolding was examined with the Luciferase Assay System. (0084)

Notes: Reporter studies were performed in primary rat ventricular myocytes, HepG2 cells and SK-N-MC cells using the pGL3 Basic Vector. All luciferase values were normalized to β-galactosidase activity provided by the pSV-β-Galactosidase Control Vector. PCR-generated deletion mutants of promoter constructs were subcloned into the pGEM^®-T Vector and sequenced. (1491)

Notes: Luciferase studies were performed in HL-60 cells. Experimental constructs were prepared in pGL3 Basic Vector and luciferase activities determined with the Luciferase Assay System. (0796)

Notes: Authors performed luciferase assays using the pGL3 Basic Vector in Calu-6 cells. (1151)

Notes: Luciferase studies were performed in HepG2 cells using constructs prepared in the pGL3 Basic Vector. (1137)

Notes: Luciferase studies were performed in NIH3T3 cells expressing 20,000 muscarinic acetylcholine receptors per cell using constructs prepared in the pGL3 Promoter Vector. (1276)

Notes: Studies were performed in the neuroblastoma cells lines SY5Y and SK-N-BE and the human rhabdomyosarcoma cell line TE671. All reporter activities were reported as fold-stimulation over pGL3 Basic Vector basal expression. (1570)

Notes: The Dual-Luciferase^® Reporter Assay System was used to study transfections of IMR32 and NIH3T3 cells with firefly luciferase vector (pGL3 Basic) constructs. Transfections were controlled with co-transfected pRL-CMV Vector. Tth DNA Polymerase was used as a high temperature reverse transcriptase. SP6 RNA Polymerase was used to produce probes for RNase protection assays. (0558)