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Sort population expressing nls-LacZ

LacZ FACS FluoReporter flow cytometry nls-LacZ

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#1 clementine

clementine

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Posted 21 September 2012 - 10:46 AM

Hello,

I've been trying to sort a cell population (from an in vivo cell suspension) that expresses LacZ with a nuclear localization signal (nls) under the promoter of a marker gene.
Initially I tried FluoReporter kit (Attached File  FluoReporterKit.pdf   132.59KB   146 downloads) from Invitrogen, but I suspect that the fluorogenic B-galactosidase substrate from the kit is not making it to the nuclei with the hypotonic shock. This protocol would not work in fixed cells since the substrate will diffuse without giving any signal (I tried that).

Would anybody have any suggestion (other kit, procedure)?

Thank you!





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