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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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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web
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|
CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web