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GFP tagged protein expression detection

GFP Expression protein

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#1 Inbox

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Excellent

Posted 15 December 2011 - 11:44 PM

I want to know whether it is possible to differentiate cell containing one GFP linked protein expressing plasmid from two GFP linked protein expressing plasmid just by FACS.
Thanx.

#2 Rsm

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Posted 16 December 2011 - 02:35 AM

You mean if you have twice the signal intensity? Which is usually not the case, but that's a different story... I think it is rather difficult to do so. Usually you have a broad signal intensity peak, spanning half a log, due to differences in promoter activity between cells or copy number of plasmids taken up. A twofold increase would be masked by this. To differentiate two populations clearly, you'd need a 10fold difference in signal strength. There are ways however to get a more narrow peak.
I think if you have time, go ahead and try it out. And let us know how it worked.

Edited by Rsm, 16 December 2011 - 02:35 AM.

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