Stable transfected Saos-2 cells (empty vector, wtp53, p53R248Q, and p53R273C) were seeded at 1 × 105 cells in six-wells plates and then transiently transfected with HER2 promoter vector, pNeuLite (Addgene Inc., Cambridge MA), using Lipofectamine 2000 reagent (Invitrogen, CA) as described above. After 24 h, cells were lysed and samples probed with Dual-Glo®Luciferase Assay System (Promega, Madison WI) essentially as described in [18 (link)]. Assays were read with the Fluoroskan Ascent FL (Labsystems, Perú), 0.5 s integration/ well. Expression induction was calculated as the relative light units obtained for the Saos-2 cells expressing mtp53/average relative light units of the Saos-2 cells expressing empty vector, normalized with renilla relative light units. Results were plotted against the log of the compound concentration, using Prism 6 Software (GraphPad, San Diego CA).
Pneulite
Manufactured by Addgene
The PNeuLite is a compact and portable compressed air system designed for laboratory applications. It generates a reliable supply of clean, dry compressed air for various laboratory equipment and processes. The core function of the PNeuLite is to provide a consistent and controlled air pressure source for laboratory use.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Dual glo luciferase assay system, by Promega (1 mentions)
Prism 6, by GraphPad (1 mentions)
Infinite m200 pro microplate reader, by Tecan (1 mentions)
Galacto light plus β galactosidase reporter gene assay system, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Luciferase assay system, by Promega (1 mentions)
2 protocols using pneulite
1
HER2 Promoter Activity Assay
2
HEK293 Dual Luciferase Assay
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HEK293 cells were plated in 6 well plate and transfected with 1 µg of pNeuLite (Addgene) alone or in combination with 0.5 µg of pcDNA3.1-flag-ELF3WT (provided from Dr. Seung Bae Rho, National Cancer Center, Republic of Korea) and p3Xflag-myc-CMV26-MED23 mutant plasmids as indicated. For the normalization of the luciferase activity of each sample, 0.5 µg of β-galactosidase expression plasmid (provided from Dr. Eun-Sook Hwang, Ewha Womans University, Republic of Korea) was also added in every sample groups. All of the transfections were conducted using Lipofectamine® 2000 Transfection Reagent (Invitrogen, USA). After 24 h, firefly luciferase and β-galactosidase activities were evaluated with the Infinite M200 PRO Microplate reader (Tecan Group ltd., Switzerland) using the Luciferase Assay System (Promega) and Galacto-Light Plus β-Galactosidase Reporter Gene Assay System (Invitrogen), respectively, according to the manufacturers’ protocols.
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