Intranuclear staining of proteins in heterogeneous cell populations and verification of nuclear localization by flow cytometric analysis

We describe techniques to detect nuclear transcription factors in thymocyte subsets using flow cytometry. We have adapted a method that minimizes autofluorescence of fixed cells, thereby allowing the detection of proteins expressed at low levels. An accompanying method in which the cytoplasm is removed from stained cells allows the confirmation of nuclear localization. These methods combine to provide a sensitive alternative approach for detecting nuclear proteins within heterogeneous cell populations. (C) 2003 Elsevier B.V. All rights reserved.