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

J Immunol Methods. 2003 Aug;279(1-2):193-8. doi: 10.1016/s0022-1759(03)00184-4.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antibodies / immunology
  • Cell Nucleus / immunology
  • Cell Nucleus / metabolism*
  • DNA-Binding Proteins / immunology
  • Early Growth Response Protein 1
  • Flow Cytometry / methods*
  • Immediate-Early Proteins*
  • Mice
  • Nuclear Proteins / immunology
  • Proto-Oncogene Proteins / immunology
  • Proto-Oncogene Proteins c-bcl-2
  • Staining and Labeling / methods*
  • Thymus Gland / cytology
  • Thymus Gland / immunology
  • Transcription Factors / immunology

Substances

  • Antibodies
  • DNA-Binding Proteins
  • Early Growth Response Protein 1
  • Egr1 protein, mouse
  • Immediate-Early Proteins
  • Nuclear Proteins
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • Transcription Factors
  • Bcl2 protein, mouse