Conditioned media from treated and untreated macrophages was collected, spun down, and filtered. This media was stored at −80˚C until use. MDA-MB231.luc (MDA-MB231 cells stably expressing luciferase) were plated in complete RPMI media in a 96 well white walled plate at a concentration of 2,000 cells per well. Conditioned media was added to the wells to make it a final concentration of 50% conditioned media. Luciferase activity was measured on day 1, 3 and 4, to assess cell growth over time in the various conditions. Luc-Screen Extended-Glow Luciferase Reporter Gene Assay system (ThermoFisher #T1035) was used according to the manufacturer’s guidelines to measure luciferase on a TECAN Infinite M200 Pro plate reader.
Luc screen extended glow luciferase reporter gene assay system
Manufactured by Thermo Fisher Scientific
The Luc-Screen Extended-Glow Luciferase Reporter Gene Assay system is a laboratory equipment product designed to detect and quantify the expression of luciferase reporter genes. It provides a bioluminescent assay solution for measuring gene expression in various experimental models.
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Lab products found in correlation
2 protocols using luc screen extended glow luciferase reporter gene assay system
1
Macrophage-Conditioned Media Effect on Tumor Cell Growth
2
Perilipin 5 3'-UTR Luciferase Assay
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The potential miR-4284 binding site at the 3'-untranslated region (3'-UTR) of Perilipin 5 (PLIN5) was inserted at the downstream of the firefly luciferase open reading frame (ORF) driven by a CMV promoter in a pCMV-Firefly Luc vector (16156; Thermo Fisher Scientific). Luciferase construct containing mutated 3'-UTR was prepared with a standard overlap-extension protocol. To determine the luciferase activity, HT-29 or SW480 cells in a 24-well plate (5 × 10 4 per well) were co-transfected with 200 ng of wild-type or mutant luciferase constructs, and 400 ng of hsa-miR-4284 or negative control, using Lipofectamine 2000 (Invitrogen). After 48 h, cells were harvested and assessed using the Luc-Screen Extended-Glow Luciferase Reporter Gene Assay System (Thermo Fisher Scientific), following provided protocols.
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