PKH and intracellular staining

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I was wondering if there is anyone out there who can help me.

We have used the membrane binding dye PKH26 to follow the division of
keratinocytes with some success. Recently we have wanted to combine this
with some intracellular staining for differentiation markers.

An initial experiment using saponin to permeabilise the cells was a tad
disappointing in it appeared to abolish the membrane staining (the
unpermeabilsed control was very bright). Has anyone had any ecxperience
of this? Are there any vital refernces that I am missing?

Thanks in advance

Derek

Derek Davies
Imperial Cancer Research Fund
London, UK
http://www.icnet.uk/axp/facs/davies/index.html

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CD ROM Vol 2 was produced by 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:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu