HeLa pLuc705 cells

Culture HeLa pLuc705 cells (may be purchased from Gene Tools,USA) as exponentially growing subconfluent monolayers in DMEM (Gibco) supplemented with... 

Wash exponentially growing HeLa pLuc705 cells with PBS to remove cell culture medium, treat with trypsin-EDTA for 5 min, centrifuge at 900 x g at 4°C for 5 min, wash twice with PBS, centrifuge again, resuspend in D-MEM, plate on 24-well plates (1.75 x 105 cells/well), and culture overnight. 

Detach exponentially growing HeLa pLuc705 cells (3.5 x 105) with trypsin (0.05%)-EDTA-4Na (0.35 mAf), centrifuge at 900 x g for 5 min, resuspend in 2 mL OptiMEM, and incubate with the fluorochrome-labeled conjugates (between 1 and 2.5 pM) for 30-120 min. 

Detach exponentially growing HeLa pLuc705 cells with trypsin-EDTA, plate on 24-well plates (1.75 x 105 cells/well), and culture overnight. 

HeLa pLuc705 cells are stably transfected by a luciferase construction allowing the quantitative assessment of PNA-peptide conjugates nuclear delivery and biological activity (Fig. 1). Cells should not be passaged more than ten times and should be checked routinely (—one to two times per month) for the absence of mycoplasma contamination... 

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See also in sourсe #XX -- [ ]

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