4×105 TOLEDO cells were electroporated and resuspended in buffer R (Life Technologies) using the Neon Electroporation Transfection System (Life Technologies) (10 μL tip; 1350 V; 1 pulse, 40 ms). In all transfections, 390 ng of the recombinant CD55-prom-enh-Nluc were combined with indicated amounts of pMT2T-BCL-6 or 100 ng of pMT2T-BCL-6-ZF/pMT2T-BCL-6-∆ZF, a Firefly luciferase (FFLuc) encoding vector (pGL4.13, Promega) to control for electroporation efficiency, and pUC19 vector (NEB) for a total of 500 ng. Electroporated cells (4 × 105 cells/mL) were cultured in IMDM/10% FBS without antibiotics, and luciferase levels measured 48 h post-transfection using the Nano-Glo Dual-Luciferase Reporter Assay System (Promega). Reporter activity was reported as the ratio between Nanoluc and Firefly (transfection control) luciferase readings in multiple technical replicates across n>3 experimental replicates. Statistical significance was calculated based on the average of technical replicates in each experiment, using a Student’s t-test (unpaired, two-tailed).
Firefly luciferase ffluc encoding vector
Manufactured by Promega
The Firefly luciferase (FFLuc) encoding vector is a laboratory tool used to express the firefly luciferase protein in cells or organisms. The vector contains the genetic sequence for the luciferase enzyme, which catalyzes a bioluminescent reaction when provided with the substrate luciferin.
Automatically generated - may contain errors
2 protocols using firefly luciferase ffluc encoding vector
1
Nanoluc Luciferase Assay for BCL-6 Regulation
2
Nanoluc Luciferase Assay for BCL-6 Regulation
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4×105 TOLEDO cells were electroporated and resuspended in buffer R (Life Technologies) using the Neon Electroporation Transfection System (Life Technologies) (10 μL tip; 1350 V; 1 pulse, 40 ms). In all transfections, 390 ng of the recombinant CD55-prom-enh-Nluc were combined with indicated amounts of pMT2T-BCL-6 or 100 ng of pMT2T-BCL-6-ZF/pMT2T-BCL-6-∆ZF, a Firefly luciferase (FFLuc) encoding vector (pGL4.13, Promega) to control for electroporation efficiency, and pUC19 vector (NEB) for a total of 500 ng. Electroporated cells (4 × 105 cells/mL) were cultured in IMDM/10% FBS without antibiotics, and luciferase levels measured 48 h post-transfection using the Nano-Glo Dual-Luciferase Reporter Assay System (Promega). Reporter activity was reported as the ratio between Nanoluc and Firefly (transfection control) luciferase readings in multiple technical replicates across n>3 experimental replicates. Statistical significance was calculated based on the average of technical replicates in each experiment, using a Student’s t-test (unpaired, two-tailed).
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