Approximately 90-95% of cell surface mabs I have tested "work" on cell fixed with 4% PFA and permeabilized with 0.1% saponin. It all depends if the epitope that mAb recognizes is denatured by PFA. We found the Caltag anti-human CD14 "works" on fixed and permeabilized samples whereas the PharMingen clone did not. This is not a reflection of the quality of the products, just the luck of the draw in regards to epitope specificity. The PharMingen catalog has tables as far as if specific clones have been tested to "work" with paraffin (formalin fixed) sections. If it does you can assume that it will work on paraformaldehyde fixed specimens as well. An added confounder is that monocyte CD14 is downregulated after LPS activation, making ID of monocytes difficult. Calman > ---------- > From: Bratislav Janjic > Reply To: royal011@visto.com > Sent: Monday, May 29, 2000 9:30 PM > To: Cytometry Mailing List > Subject: Surface and intracellular staining > > > Dear fellows, > > Is there any possibility to perform simultaneous labeling (I mean, in the > same sample) > of surface and intracellular antigens? Is it possible, for example, to > perform labeling > of monocytes with CD14, and after that, to permebilise them and label > intracellular > antigens (for example caspases)? Does the cell permeabilising affect > labeling of > surface antigen? If somebody has experience with similar procedures, > please contact me. > > Thanks in advance. > > Bratislav Janjic, BS > Department of Pathology > University of Pittsburgh > > > > __________________________________________________________________________ > _ > Visit http://www.visto.com/info, your free web-based communications > center. > Visto.com. Life on the Dot. >
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