Image - Barts and The London logo and link to home page Image - divider Image - divider
  Institute of Cell and Molecular Science
Flow Cytometry Core Facility equipment, images used with permission by BD
  Flow Cytometry Core Facility
  link Home link Instruments available link Flow cytometry link Uses of flow cytometry link Flow sorting link Links link Contacts

Cytoplasmic / nuclear antigen analysis


To detect intracellular antigens it is necessary to permeabilise the cell. Many reagents are available commercially to facilitate this.

There is a wide variety of different types of intracellular antigen that can be detected by flow cytometry, including:-

Cytokines -Th1Th2 responses

Lymphocytes and splenocytes involved in inflammatory or allergic responses produce a preponderance of IFNgamma or IL-4 respectively. KO murine splenocytes Th1Th2 responses were compared to wild type splenocytes. The KO showed an increased capacity to produce IFNgamma compared to the wild type response whilst IL-2 and IL-4 levels remained the same as the wild type.

Detection of intracellular IL-17

Interleukins (e.g. IL1-17) can be measured intracellularly in lymphocytes after stimulation typically with PMA/ionomycin and blocking the Golgi apparatus with the antibiotic Brefeldin A or Monensin, see protocol.

Transcription factors

Foxp3 the self-reactive transcription factor, the master regulator involved in lymphocyte development and function of regulatory T cells can be measured intracellularly by flow cytometry, see protocol.


The intracellular detection of cyclins can be combined with cell cycle analysis, see cell cycle analysis section.

Apoptotic factors

The pro-apoptotic factor, bak undergoes a conformational change when activated during apoptosis involving the mitochondria. The degree of bak activation can be determined intra- cellularly by use of mcab that detects the activated form of bak, see protocol.


Th1Th2 phenotyping of murine splenocytes

IL-17 expression in murine peripheral blood lymphocytes

Foxp3 expression in murine peripheral blood lymphocytes

Bak activation in human fibroscarcoma

by Gary Warnes. © Queen Mary, University of London 2007
Institute of Cell and Molecular Science, The Blizard Building, Barts and The London School of Medicine and Dentistry, 4 Newark Street, London E1 2AT, UK Tel: +44 (0)20 7882 2483, Fax: +44 (0)20 7882 2200