HeLa cells were grown in DMEM (HyClone) supplemented with 10% Fetal Bovine Serum (VWR) and 1× Penicillin/Streptomycin (HyClone) in 24 well plates. When the cells reached 80% confluence, co-transfection of PHOX2B-pcDNA3.1 (245ng), DBH-pGL3-basic (245ng, courtesy of Dr. Sucheta Vaingankar University of California, San Diego), and pRL-TK (24.5ng, Promega) was performed using Lipofectamine 3000 (ThermoFisher) following the manufacturer’s suggested protocol. Forty eight hours after transfection, cell lysates were collected in 100ul of 1× Passive Lysis Buffer (Promega). Firefly and Renilla luciferase activity was detected using the Dual-Luciferase Reporter Assay System (E1910, Promega) following the manufacturer’s suggested protocol. Luciferase activity was read using a Promega GlowMax 96 well luminometer (Promega).
Glowmax 96 well luminometer
Manufactured by Promega
The Promega GlowMax 96 well luminometer is a compact and efficient instrument designed for high-throughput luminescence detection. It features a 96-well format and provides accurate and reliable measurements of luminescent signals, making it suitable for a variety of assays that utilize luminescent detection methods.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Avantor (2 mentions)
Dual luciferase reporter assay system, by Promega (2 mentions)
Prl tk, by Promega (2 mentions)
Passive lysis buffer (plb), by Promega (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Promega (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
2 protocols using glowmax 96 well luminometer
1
Dual-Luciferase Reporter Assay for PHOX2B and DBH
2
Transcriptional Regulation of FOXP1 in HEK293 Cells
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HEK 293 cells were grown in DMEM (Corning) supplemented with 10% Fetal Bovine Serum (VWR) and 1% Penicillin/Streptomycin (HyClone) in 24 well plates. When the cells reached 80% confluence, co-transfection of pcDNA3.1-FOXP1 (250 ng), DBH-pGL3-basic (250 ng), and pRL-TK (25 ng, Promega) was done using Lipofectamine 2000 (ThermoFisher) following the manufacturer’s suggested protocol. After 24 hours, fresh DMEM was added and 24 hours later the cells were lysed using 1X Passive lysis buffer (Promega). Firefly and Renilla luciferase activity was detected using the Dual-Luciferase Reporter Assay System (Promega) following the manufacturer’s suggested protocol. Luciferase activity readings were acquired on biological triplicates and technical triplicates using a Promega GlowMax 96 well luminometer (Promega). 1-way ANOVA with Fisher’s LSD test were used for statistical analysis.
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