Cell lines C30, C30-FRβ, THP1, MV411, and HL60 were stably transduced with GFP-2A-firefly luciferase (fLuc) lentiviral vector. 2×104 fLuc-targets were cultured in white 96-well plates with CAR+ T-cells at 5:1, 1:1, and 1:5 E:T ratios in triplicate. After overnight co-culture, residual luciferase activity was measured using the Extended-Glow Luciferase Reporter Gene Assay System (Life Technologies). Percent lysis was calculated as follows [100− ((Average Signal T-cell Treated Wells)/(Average Signal Untreated Target Wells)×100)].
Extended glow luciferase reporter gene assay system
Manufactured by Thermo Fisher Scientific
The Extended-Glow Luciferase Reporter Gene Assay System is a laboratory equipment product designed to measure luciferase reporter gene activity. The system utilizes a luciferase enzyme that produces a prolonged luminescent signal, allowing for extended monitoring and quantification of gene expression.
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2 protocols using extended glow luciferase reporter gene assay system
1
Cytotoxic Activity Assay for CAR T-cells
2
Cytotoxic Activity Assay for CAR T-cells
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Cell lines C30, C30-FRβ, THP1, MV411, and HL60 were stably transduced with GFP-2A-firefly luciferase (fLuc) lentiviral vector. 2×104 fLuc-targets were cultured in white 96-well plates with CAR+ T-cells at 5:1, 1:1, and 1:5 E:T ratios in triplicate. After overnight co-culture, residual luciferase activity was measured using the Extended-Glow Luciferase Reporter Gene Assay System (Life Technologies). Percent lysis was calculated as follows [100− ((Average Signal T-cell Treated Wells)/(Average Signal Untreated Target Wells)×100)].
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