I think I know the artifact you are talking about. With intrcellular staining for anything- the non-specific staining is a big problem. If your antibodies have different fluorochromes you can have one appear to stain more intensely with one antibody as if it is monoclonal but it is just non-specific. We have also found that some polyclonal antibodies are stickier than others and the nonspecific staining is not equal. This is exasperated when you have intracellular staining. I usually use intracellular only for plasmacytic malignancies and the staining is so bright when it is positive you can tell. Maryalice >Hello Flowers: >I wonder if any one has some experience with the kappa /lambda preparation on >permeabilized cells ( Intraprep) for determinig cytoplasmic >expression of the light >chains and hence clonality ( restriction) in B cells. It seems that >occasionally, >a preparation artifact occurs where only one light chain is >expressed in otherwise >polyclonal populations of B cells. Commonly we use the Intraprep >when surface labeling >is very dim or negative. The common combination is >CD45/CD19/kappa/lambda. The CD19 >is ususally the fourth color-PC5 >I would appreciate some comments on this problem >Thanks. -- Maryalice Stetler-Stevenson Director Flow Cytometry Unit Laboratory of Pathology, NCI, NIH "Learn the rules so you know how to break them properly." The Dalai Lama
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