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A few years ago we did a lot of work labelling a ras oncogene product p21 with afluorescein conjugated antibody. We found that fixation produced quite a bit ofbackground. We ultimately used lysolecithin to permeabilize the cells, which did not result in
much background, and got nice histograms with many intact cells. We also have used
electroporation to open pores in the plasma membrane, allowing antibody to pass
with cells still viable. Worked great! I would suggest conjugating your antibody
Debbie Berglund
Microbiology
Montana State University
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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