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Expression systems: Difference between revisions
→Human Embryonic Kidney (HEK) 293 cultured cells
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===Human Embryonic Kidney (HEK) 293 cultured cells === | ===Human Embryonic Kidney (HEK) 293 cultured cells === | ||
Commonly knows as HEK293 cells they are actually an excellent protein source, especially for secreted proteins. Contrary to urban legend, they are actaully significantly cheaper than instect cells to culture. You need standard cell culture equipment that in most cases is available in any biology department. Quite easy to scale up the volume still using attached cells, in special plastic bottles that present a large surface area: thats most useful especially for secreted proteins. There are two main ways to express your gene in HEK293 cells: | |||
*[http://www.ncbi.nlm.nih.gov/pubmed/17001101?ordinalpos=6&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum Transient transfection]. | *[http://www.ncbi.nlm.nih.gov/pubmed/17001101?ordinalpos=6&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum Transient transfection]. | ||
*[http://www.invitrogen.com/content.cfm?pageid=3375 Flp-In | *[http://www.invitrogen.com/content.cfm?pageid=3375 Flp-In ] to generate stable cell lines. | ||
Producing intracellular proteins in HEK293 cells requires to grow the cells in suspension which can be a bit more of an art and requires a CO2 shaking incubator. | |||
=Other systems= | =Other systems= | ||