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> We are trying a saponin permeabilization technique - to measure
> intracytoplasmic cytokines and other proteins (p24 etc). We are
> initially fixing with 4% formaldehyde and then coming in with saponin
> and then labelling with our Ab.
Final conc 4% or 2%? Our protocol uses equal volumes of cells and 4%, i.e.
2% final, slightly hypertonic. Duration of fix and temperature can also be
critical.
*******************************************************************
*Peter Openshaw p.openshaw@ic.ac.uk *
*Respiratory Medicine Tel:+44 171 723 1252 x 5781/6 *
*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