Notes: To study the potential interactions among the five SALMs (synaptic adhesion-like molecules) family members, the cDNAs of the five SALMs were subcloned and then transiently transfected in various combinations of two to five SALMs into HEK293 cells. The cells were transfected using calcium phosphate precipitation with an equimolar amount of the pAdVAntage™ Vector to enhance protein production. Interactions of the expressed SALMs were analyzed by immunoprecipitation experiments. (3754)

Notes: This study looked at how OX40, a member of the TNFR superfamily, affected the differentiation of CD8 T cells with an OX40 agonist. Recombinant lentiviruses were generated by cotransfection of a vector plasmid, virus-packaging plasmid, envelope plasmid and the helper pAdVAntage™ Vector into HEK 293 cells. The supernatant was collected and used to infect tumor cells which were injected into mice. (3753)

Notes: The authors used reporter assays and coimmunoprecipitation experiments to compare the zebrafish (Danio rerio) Toll-IL-1R-containing adaptor molecule 1 (TICAM1) activation of NF-κB and zebrafish type I IFN to mammalian TICAM1 activation. 293H and ZFL (zebrafish liver) cells were cotransfected with 400 ng of TICAM1 construct (mouse, zebrafish or a deletion construct), 400 ng of a reporter construct (e.g., a zebrafish IFN promoter cloned into the pGL3-Basic Vector) and 10 ng of pRL-CMV Vector, an internal control to normalize data. After 24 hours for 293H cells and 48 hours for ZFL cells, the transfected cells were lysed, and luciferase activity was measured using the Dual-Luciferase^® Reporter Assay System. For coimmunoprecipitation experiments, 293H cells were cotransfected with a total of 8µg of plasmids (3µg of zebrafish or mouse TICAM1 construct, 3µg of interacting protein construct, 1µg of the pAdVAntage™ Vector, and 1µg of antiapoptotic protein p35 construct). Forty-eight hours posttransfection, the cells were lysed, the protein bound to affinity resins and analyzed by Western blot. (3755)

Notes: To examine the interaction between endogenous cellular prion protein (PrP^C) and misfolded, disease-associated (PrP^Sc), hybrid antibodies were created using domains from PrP^C grafted onto the b12 protein. These antibodies were expressed using 30µg of plasmid to transiently transfect a suspension of 293 human embryonic kidney cells in a serum-free medium. To enhance antibody expression, 5µg of the pAdVAntage™ Vector was co-transfected. The cells were harvested 48–72 hours post-transfection, the supernatant centrifuged and the antibody purified over a protein A-Sepharose column. (3573)

Notes: To determine if fusion of the HIV virion to maturing dendritic cells (DCs) was the limiting step in HIV replication in DCs, β-lactamase-Vpr (BlaM-Vpr) chimeric virions were produced for use in a HIV virion fusion assay. To generate the virion chimeras, 293T cells were transfected with pNL4-3 or 81A proviral DNA, pCMV-BlaM-Vpr and pAdVAntage™ vectors. The virions were pseudotyped with various HIV env protein constructs. Forty-eight hours post-transfection, the virus-containing supernatant was harvested by centrifugation and normalized to p24 Gag content as measured by ELISA. (3495)

Notes: To produce peptide epitope-embedded antibodies, 1.2 × 10⁷ 293 EBNA cells were seeded in 150mM tissue culture plates. The following day, each dish was transfected using calcium chloride complexed with 16µg of the IgG expression plasmid along with 4µg of pAdVAntage™ Vector and 800ng of pEGFP-1. After 24 hours, the medium was changed to serum-free medium. After an additional 24 hours, 2.5 ml of 0.5 M HEPES/20% glucose was added. Cells were incubated 4 days, and the antibody in the medium supernatant was purified by protein A chromatography. (3494)

Notes: Researchers demonstrated an increase in reporter protein activity when the pAdVAntage™ Vector was cotransfected into NIH3T3 cells with pCLMFG, a viral vector, and pCL-Ampho, a packaging vector. A direct correlation was observed between reporter activity and increasing amounts of pAdVAntage™ Vector in transfections. (3015)

Notes: The pAdVAntage™ Vector was cotransfected with a pCAT-control vector into 293 cells. A CAT assay was performed 48 hours post-transfection and results were compared to those from cells cotransfected with a pCAT-control vector and an experimental vector expressing influenza A virus NS1 protein.  In all transfections the total amount of DNA used was 2.5μg. (2753)

Notes: The pAdVantage™ Vector was contransfected with CAT reporter vector, CMV-driven expression vector and a LacZ control vector to increase transient expression levels in HeLa cells (0822)

Notes: The pAdVAntage™ Vector was cotransfected with an expression vector containing the alpha-2 subunit of the calcium channel. The pAdVAntage™ Vector did not change the pharmacological characteristics of the alpha-2 subunit. (1378)

Notes: The pAdVAntage™ Vector was cotransfected with a green fluorescent protein construct into 293 cells. (1977)