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>
>
> We are trying to determine if COS cells are expressing a particular
> protein after transient transfection. The protein is designed to be
> secreted but I am hoping we can find enough of it in the cytoplasm to tell
> us the experiment worked before we do a Western blot on the culture
> supernatant.We have directly PERCP conjugated mouse monoclonal antibody to
> this protein. Do you have any recommended protocol for permeablization,
> staining, washing etc.? Since these cells are quite large, is there
> anything we should be doing with the FSC and SSC, other than using linear
> amplification? We are using a BD FACSCAN with LYSIS software.
>
> Happy Holidays.
>
> Howard
>
*******************************************************************
*Peter Openshaw (p.openshaw@ic.ac.uk) Direct:0171 594 3854 *
*Respiratory Medicine Switch:+44 171 723 1252 x 3854/3 *
*St. Mary's Hosp. Med. School Fax:+44 171 724 7349 *
*Imperial College, Norfolk Place, London W2 1PG, UK *
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