Notes: The Dual-Luciferase™ Reporter System was used to quantitate the presenilin promoter activity in Neuro2a neuroblastoma cells, mouse P19 embryonal carcinoma cells and NIH 3T3 cells. Studies were also performed in P19 cells treated with retinoic acid to acquire a neuron-like phenotype and P19 cells treated with dimethyl sulfoxide to acquire a muscle-like phenotype. The presenilin promoter functioned best in the Neuro2a and neuron-like P19 cells. 5´-RACE products from mouse brain RNA were purified with the Wizard^® PCR Preps System and cloned into the pGEM-T Vector . The cloned amplimers were sequenced and used as a template for amplification to produce truncation mutants to assess promoter activity. (1590)

Notes: The PolyATtract^® System 1000 was used to isolate polyA+ RNA directly from newborn mouse brains. The RNA was used in RT-PCR for detection of specific TrkB variants. Taq DNA Polymerase and the pGEM^®-T Vector System were used to produce and subclone novel TrkB variants that differ in the extracellular domain. The CellTiter 96^® Non-Radioactive Cell Proliferation Assay was performed on NIH 3T3 cells transfected with the TrkB receptors and challenged with BDNF, NT3 and NT4/5. (0615)

Notes: Sixty patients were screened for autoantibodies to IA-2 (a protein tyrosine phosphatase-like molecule), IA-2ic (the intracellular fragment of IA-2) and GAD65 (glutamic acid decarboxylase) using RIAs. Full-length human cDNAs were cloned into the pGEM^®-4Z Vector and expressed in the TNT^® Coupled Reticulocyte Lysate System in the presence of [³⁵S]methionine. Incorporated radiolabel was determined by precipitation and scintillation counting. Immunoprecipitation was performed with 5µl of serum and counted on a multiwell counter. Antibodies to IA-2 or GAD65 were detected in 87% of newly diagnosed type 1 diabetic patients and in all 56 prediabetic samples from 11 twins. The frequencies of antibodies to IA-2ic were similar in type 1 diabetics. (1763)

Notes: Promega's Tth DNA Polymerase, pGEM^®-T Vector System and Riboprobe^® System were used in this study. (2002)

Notes: The pGEM^®-T Vector System was used in this study. (2194)

Notes: PCR products were cloned into the pGEM^®-T Vector System. (0082)

Notes: The system was used to clone the amplimers generated by RT-PCR. Sequencing of the clones was performed in the vector. (1569)

Notes: The pGEM^®-T Vector System was used in this study. (2005)

Notes: The pGEM^®-T Vector System was used in this study. (1970)

Notes: The pGEM^®-T Vector System was used to clone PCR products amplified from a tobacco cDNA library. (1968)

Notes: The pGEM^®-T Vector System was used in this study to clone amplified cDNAs. (1973)

Notes: The pGEM^®-T Vector System was used in this study. (2193)

Notes: The pGEM^®-T Vector System was used in this study. (1967)

Notes: HA and Myc epitope tags were introduced at the C-terminus of Cdc1 and Cdc27 by oligo-directed mutagenesis. Cdc1tag and Cdc27tag were cloned into the pGEM^®-4Z Vector and expressed in the TNT^® T7 Coupled Reticulocyte Lysate System ± [³⁵S]methionine. In GST coimmunoprecipitation assays, Cdc1 interacts with GST-Pol3, and the GST-Pol3 binds preferentially to a truncated Cdc1 translation product. To confirm an interaction observed between Cdc1 and Cdc27, the TNT^®-expressed proteins were mixed together and co-immunoprecipitated using the anti-Cdc27 antibodies Using EGS (ethylene glycol-bis-succinimidylsuccinate) labeled, in vitro translated Cdc1 and Cdc27 were cross-linked. (1771)

Notes: The pGEM^®-7zf(-) vector was used for subcloning cDNAs. The pCAT^®3-Control Vector was used as a control in studies of CAT reporter constructs in CV1 and HeLa cells. GST and GST-YY1 fusion proteins were produced using the TNT^® T7 Coupled Reticulocyte Lysate System. (0141)

Notes: A 3,375 bp fragment was cloned in the pGEM^®-T Vector System. (0223)

Notes: Full-length and fragments of [35S]methionine-labeled U1 snRNP-A protein were produced using the TNT® Coupled Reticulocyte Lysate System. The proteins were used in RNA-binding assays to the U1 RNA, SV40 RNA, pGEM® RNA (negative control) and triple mutant of SV40 sequences. The results show that the U1 snRNP-A is able to coprecipitate the U1 RNA and SV40 RNA, but a ten-fold reduction in precipitation of the triple mutant is seen. Both U1 fragments could bind the mutant but at greatly reduced levels. One U1 fragment precipitated both the U1RNA and the SV40 RNA; the other U1 fragment only precipitated the SV40 RNA. (1837)

Notes: Utilize protein fusions within pGEMEX^®-1 Vector and partially purify protein products using MAb against P2 protein product. (0735)