https://eln.excellgen.com/index.php?action=history&feed=atom&title=293FE_Cell_Line 2026-10-18T10:13:40Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=293FE_Cell_Line&diff=5751&oldid=prev 2012-08-22T22:54:09Z

<p></p> <p><b>New page</b></p><div>HEK293SF-3F6 (ATCC Accession No. CRL-12585) stably expressing the EBNA1 protein. The SFE cell line was developed by transfecting the 293SF-3F6 cell line with the pIRES-neo vector encoding the full-length EBNA1 protein and isolating and amplifying geneticin-resistant clones so obtained. The isolated clones were then tested for transient expression of the SEAP gene. A clone (clone 41) showing the highest expression levels was deposited under IDAC Accession No. 020502.&lt;br&gt; <br /> <br /> &lt;br&gt;<br /> <br /> The cell line was cultured in HyQSFM4TransFx293 (HyQ) (Hyclone, Logan, UT, USA)&amp;nbsp; supplemented with 5% Cell Boost 5 (CB5, Hyclone).&lt;br&gt;<br /> <br /> Pre-cultures underwent passages every 2-3 days to keep the cells in their exponential growth phase. A few hours before transfection, the cell suspension was&lt;br&gt;centrifuged (300 g for 5 min) and re-suspended in fresh medium at 5 × 10^6 cell/mL. three different DNA concentrations&lt;br&gt;(0.4, 0.6, and 0.8 μg DNA/106 cells) were tested at 37°C.&lt;br&gt;The potential of repeated transfections at three different&lt;br&gt;times (0, 48, and 96 hours post-inoculation) and two&lt;br&gt;different temperatures (34 and 37°C) were also tested. In&lt;br&gt;the repeated transfection experiments, a new amount of&lt;br&gt;polyplexes was added at 48 and 96 hours, besides the&lt;br&gt;amount added in the beginning of the experiment&lt;br&gt;&lt;br&gt;&lt;br&gt;&lt;br&gt;<br /> <br /> [[Category:293_Expression]]</div>

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