HEK293T cells were seeded into 96-well culture plates and transfected with 1 μg of each constructed the next day. 24 h post transfected, we washed the cells with PBS and treated them with lysis buffer (Pierce Firefly Luciferase Glow Assay Kit, Thermo Fisher Scientific) for 20 min. The cell lysate was then collected and spun down to remove cell debris. 15 μl of the cell lysate was transferred into a 96-well black polysterene plate (Corning Costar). 45 μl of the working buffer (D-luciferin and the Firefly Glow Assay buffer according to manufacturer’s instructions, Pierce Firefly Luciferase Glow Assay Kit, Thermo Fisher Scientific) was added to the cell lysate and we waited 10 min before acquiring signals using a microplate reader (Tecan, Männedorf, Switzerland).
96 well black polysterene plate
Manufactured by Corning
The 96-well black polystyrene plate is a lab equipment product used for various scientific applications. It is a multi-well plate made of black polystyrene material, designed with 96 individual wells arranged in a 8x12 grid format. The product's core function is to provide a standardized and well-defined container for holding and testing small volumes of liquids or samples in a high-throughput manner.
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2 protocols using 96 well black polysterene plate
1
Luciferase Assay in HEK293T Cells
2
Firefly Luciferase Assay in HEK293T Cells
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HEK293T cells were seeded into 96-well culture plates and transfected with 1μg of each constructed the next day. 24 hours post transfected, we washed the cells with PBS and treated them with lysis buffer (Pierce Firefly Luciferase Glow Assay Kit, Thermo Fisher Scientific) for 20 minutes. The cell lysate was then collected and spun down to remove cell debris. 15μl of the cell lysate was transferred into a 96-well black polysterene plate (Corning Costar). 45μl of the working buffer (D-luciferin and the Firefly Glow Assay buffer according to manufacturer's instructions, Pierce Firefly Luciferase Glow Assay Kit, Thermo Fisher Scientific) was added to the cell lysate and we waited 10 minutes before acquiring signals using a microplate reader (Tecan, Männedorf, Switzerland).
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