Reporter for macrophage (J774) transfection

tronni at my-deja.com tronni at my-deja.com
Sat Nov 13 03:31:25 EST 1999


In article <spam-0911991243290001 at macmac-2.ucsf.edu>,
  spam at 127.0.0.1 (Bernard Murray, PhD) wrote:
> In article <808orm$i0h$1 at nnrp1.deja.com>, tronni at my-deja.com wrote:
>  I would use CMV-driven Green
> > Fluorescent Protein (GFP) reporter as marker for transfection.
> > With this you can easily estimate the relative amount of transfected
> > cells by immunofluorescence or flow cytometry (with 488 nm
argon-laser).
>
> Ah, but if you have a FACS you could sort the transfected cells
> and collect the green ones (100% transfection efficiency!).
>

True, if you have access to a fast cell sorter. Those are
expensive and require a lot of experience to use. Fastest model I'm
aware of is able to sort 15,000 cells/second. For gene regulation
studies, at least, FACS sorting of J774 cells is not a feasible option
considering time and money needed. With current
level of transfection 10 million cells would give you only 100,000
cells after sorting.


>
>    Bernard
>
> --
> Bernard P. Murray, PhD
> bpmurray at cgl . ucsf . edu
> Department of Cellular & Molecular Pharmacology, UCSF
>

Tapani Ronni, PhD
UCLA School of Medicine/Howard Hughes Medical Institute



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