Notes: Protein Truncation Test (PTT) was performed by use of the TNT^® Coupled Reticulocyte Lysate System, detection was performed by incorporation of [³⁵S]- methionine/cysteine. (0277)

Notes: Th authors used Promega's TNT® Coupled Reticulocyte Lysate System and Canine Pancreatic Microsomal Membranes (CMMs) in initial characterization of an assay for processing of a chimeric APP molecule. (1373)

Notes: Several variant hepatitis B isolates were subjected to PCR to isolate the surface antigen cDNA. The PCR products were cloned and subjected to in vitro transcription/translation with the TNT^® Coupled Reticulocyte Lysate System in the presence of Canine Pancreatic Microsomal Membranes (CMMs). The amount of microsomal membranes was adjusted to give the approximate ratios of wild-type glycosylated and unglycosylated forms. The variants were then tested for immunoprecipitation with a commonly used anti-hepatitis B surface antigen pAb. Only one of the variants was immunoprecipitated. (0520)

Notes: The human protein hsPex14p as well as another protein hsPex13p and hsPex5p were cloned into the PinPoint® Xa2 vector and expressed in JM109 E. coli. The cell extracts were run on SDS PAGE gels, transferred to nitrocellulose and probed with the anti-MF3 antisera. The hsPex5p, supposedly the human PTS1 homolog, was translated in vitro with the TNT® Coupled Reticulocyte Lysate System and reacted with another blot with the same proteins and the hsPex14p bound the 35S-labeled protein. (1172)

Notes: These authors used the TNT® T7 Coupled Reticulocyte Lysate System together with Canine Pancreatic Microsomal Membranes (CMMs) to show post-translational modification of RAIG1. They also used RQ1 RNase-free DNase to treat RNA samples prior to cDNA synthesis. (1337)

Notes: The cloned 752 amino acid GC-binding factor 2 (GCF2) was expressed in vitro using the TNT® Coupled Reticulocyte Lysate System. The synthesized product migrated at ~160kDa by SDS-PAGE when the predicted molecular weight was 83kDa. Expression of the protein from E.coli also produced a protein that migrated at ~160kDa. Mass spectrum analysis confirm a mass of 83kDa. The E.coli produced protein was further characterized by gel shift analysis with the AP-2 Consensus Oligonucleotide. Recombinant GCF2, AP-2 and SP1 transcription factors were analyzed for binding to the EGF receptor promoter by footprint analysis. The GCF2 protein was expressed with a EGF Receptor promoter-CAT reporter vector in CV-1 cells and found to increase CAT activity. (0510)

Notes: cDNA synthesis was performed in a reverse transcription reaction containing single-stranded binding protein and RNasin^® Ribonuclease Inhibitor. In a protein truncation test (PTT), five overlapping segments amplified by RT-PCR of NF1 transcript were expressed by TNT^® Coupled Reticulocyte Lysate System. (1051)

Notes: The TNT® T7 Coupled Reticulocyte Lysate System was used to synthesize and label PEX12 proteins with [35S]methionine. (1360)

Notes: The authors used TNT^® Coupled Reticulocyte Lysate Systems together with Canine Pancreatic Microsomal Membranes (CMM) to perform heterologous import experiments with trypsin protection of a diatom nuclear-encoded plastid protein. (0836)

Notes: The authors used a TnT^® Coupled Reticulocyte Lysate System to transcribe and translate Src in vitro. Cell proliferation of NIH 3T3 cells that stably expressed either HA-RACK or vector only was assessed using the CellTiter 96^® Non-Radioactive Cell Proliferation Assay. (2537)

Notes: The authors used Promega's purified DNA-Dependent Protein Kinase as a substrate for in vitro phosphorylation by the protein tyrosine kinase Lyn. They also used the TNT^® Coupled Reticulocyte Lysate System to synthesize radiolabeled DNA-dependent protein kinase (DNA-PK) catalytic subunit polypeptides for protein:protein interaction studies with GST-tagged Lyn. (0863)

Notes: BAEC (bovine aortic endothelial) cells were transfected using Transfectam^® Reagent. Serum response factor (SRF) was synthesized in vitro using the TNT^® Coupled Reticulocyte Lysate System. (1070)

Notes: cDNA was generated by reverse transcription of RNA purified from lymphoblastoid cell lines. PCR amplification of the cDNA was used to introduce a 17bp consensus T7 promoter sequence and a mammalian translation-initiation sequence in frame with a unique hMLH1 or hMSH2 sequence. Resultant PCR products were translated in the TNT^® T7 Coupled Reticulocyte Lysate System. (1194)

Notes: The authors used the TNT^® T7 Coupled Reticulocyte Lysate System and Canine Pancreatic Microsomal Membranes (CMMs) to make UDP-galactose:ceramide galactosyltransferase (CGalT) to test antibodies for their ability to immunoprecipitate this protein. (0364)

Notes: The TNT^® Coupled Transcription/Translation System was used to verify the 25.3kDa size of the cloned receptor. AMV reverse transcriptase was used for first strand cDNA synthesis prior to PCR. (1520)

Notes: Full-length, human hnRNP A1 protein and truncated hnRNP (lacking the M9 domain, A1DeltaM9) were synthesized using the TNT^® Coupled Reticulocyte Lysate System. The synthesized proteins were injected directly into oocytes without purification and were assayed for nucleocytoplasmic transport. (0964)

Notes: pSV-β-Galactosidase Control Vector was used as a transfection control in F9 cells and TNT^® T7 Coupled Reticulocyte Lysate System was used to translate ATF-1 protein for use in gel shift assays. (1004)

Notes: [³⁵S]methionine-labeled RIP-140, SRC-1, TIF1 and TIF2 were produced using the TNT^® Coupled Reticulocyte Lysate System. The labeled proteins were incubated with GST or GST-AF2 receptor fusion proteins coupled to glutathione beads in the absence or presence of oestradiol or 4-hydroxytamoxifen. After overnight incubation and washing away of free protein, bound proteins were extracted and separated by SDS-PAGE. From binding studies with wild type or mutant AF2 receptors, SRC-1, TIF1 and TIF2 require Lys-366 in AF-2 for their interaction, but RIP-140 does not. (1797)

Notes: The DNA strand break repair protein encoded by the human XRCC1 gene was previously reported to interact with the DNA ligase III-alpha protein. An XRCC1 polypeptide was synthesized with [³⁵S]methionine using the TNT^® Coupled Reticulocyte Lysate System. GST-DNA ligase fusion proteins were adenylated, separated by denaturing gel electrophoresis and transferred to nitrocellulose. After renaturation, the membranes were incubated with the labeled XRCC1 polypeptide. XRCC1 formed a complex with DNA ligase III-alpha, but there was no detectable complex between XRCC1 and DNA ligase III-beta. (1770)

Notes: This article describes the use of the TNT^® T7 Quick Coupled Transcription/Translation System in a process called the ‘ARM’ technique where antibody-ribosome-mRNA (ARM) complexes are selected by protein binding. The ARM complexes are generated during translation of mRNAs that lack stop codons. The functional antibody that is still complexed with the ribosome and mRNA is selected by protein-coupled magnetic beads, and the associated mRNA is reverse-transcribed and amplified by polymerase chain reaction (RT-PCR). The PCR products are then used in subsequent cycles of the method. (2016)

Notes: Cystic fibrosis transmembrane conductance regulator (CFTR) was expressed in the TNT^® Coupled Reticulocyte Lysate System to confirm its molecular weight. Translation was performed with Canine Pancreatic Microsomal Membranes (CMMs) to verify that the protein is integrated into the membrane. Further, the electrophoretic mobility of CFTR suggests that it is glycosylated. (0570)

Notes: This article details the use of the IVEC technique in the TNT^® Coupled Reticulocyte Lysate System to screening 100,000 cDNA clones to identify gelsolin as a substrate of caspase-3 protease. (0885)

Notes: Promega's M-MLV Reverse Transcriptase, TNT^® Coupled Reticulocyte Lysate System, and Taq DNA Polymerase were used in this study. (2000)

Notes: Various portions of a 552 residue protein were cloned and expressed in BL21(DE3)pLysS using the pGEMEX®-1Vector. The resulting proteins were purified and used as immunogens. The TNT® Coupled Reticulocyte Lysate System was used for in vitro translation. (1236)

Notes: Promega's pGEM®-T Vector System, pCI Mammalian Expression Vector and TNT® Coupled Reticulocyte Lysate System were used in this study. (1951)