CHO-K1 cells were grown to 95% confluence in compete DMEM before being transfected with 1ug of luciferase or tdTomato-expressing plasmid or Doggybone. DNA was delivered by electroporation using a Nucleofector II device with cell line transfection kit T (Lonza) or using the following transfection reagents; Lipofectamine 2000 (Invitrogen), Polyfect (Qiagen) or PEI (Polysciences). Each was used according to supplier’s specification. Luciferase expression : At the appropriate juncture, media was removed, cells were lysed using luciferase lysis buffer (Promega) and frozen for 24 hours. Expression was visualized after the addition of luciferase assay substrate (Promega) and light emission was read using a FluorStar OPTIMA multilablel plate reader (BMG Labtech) and recorded as relative light units (RLU). tdTomato red fluorescent protein expression was imaged using a Nikon Eclipse TE2000-S inverted microscope under bright light and a TxRed filter set (excitation 540-580nm and emission 600-660nm) at 24 and 48 hours after transfection.
Fluorstar optima multilablel plate reader
Manufactured by BMG Labtech
The FluorStar OPTIMA is a multilabel plate reader designed for fluorescence, luminescence, and absorbance detection. It offers rapid and accurate measurements across a wide range of applications.
Automatically generated - may contain errors
Lab products found in correlation
Luciferase lysis buffer, by Promega (2 mentions)
Luciferase assay substrate, by Promega (2 mentions)
Cell line transfection kit t, by Lonza (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Polyfect, by Qiagen (2 mentions)
Polyethylenimine (pei), by Polysciences (2 mentions)
Eclipse te2000 s, by Nikon (2 mentions)
Nucleofector 2 device, by Lonza (2 mentions)
2 protocols using fluorstar optima multilablel plate reader
1
Transfection and Reporter Assay in CHO-K1 Cells
2
Transfection and Reporter Assay in CHO-K1 Cells
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CHO-K1 cells were grown to 95% confluence in compete DMEM before being transfected with 1ug of luciferase or tdTomato-expressing plasmid or Doggybone. DNA was delivered by electroporation using a Nucleofector II device with cell line transfection kit T (Lonza) or using the following transfection reagents; Lipofectamine 2000 (Invitrogen), Polyfect (Qiagen) or PEI (Polysciences). Each was used according to supplier’s specification. Luciferase expression : At the appropriate juncture, media was removed, cells were lysed using luciferase lysis buffer (Promega) and frozen for 24 hours. Expression was visualized after the addition of luciferase assay substrate (Promega) and light emission was read using a FluorStar OPTIMA multilablel plate reader (BMG Labtech) and recorded as relative light units (RLU). tdTomato red fluorescent protein expression was imaged using a Nikon Eclipse TE2000-S inverted microscope under bright light and a TxRed filter set (excitation 540-580nm and emission 600-660nm) at 24 and 48 hours after transfection.
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