HomeImmunophenotyping of murine lymphocytes, splenocytes and thymocytes at ICMS includes the measurement of CD25 (IL-2 receptor) on lymphocytes in conjunction with CD3-FITC and CD4-APC labelling. These cells can be then further investigated for intracellular antigens such as IL-17 and foxp3. (see cytoplasmic antigen analysis).
Murine whole blood was analysed for endothelial cell stem cells by labelling with negative selection cocktail-FITC (CD45, B220, etc), CD31-PE, Sca-1-PE-Cy7 and c-kit-APC and back-gating onto FSC vs SSC after doublet discrimination, see figure for identification of endotheilial stem cells and control panel.
A mixed population of murine thymocytes consisting of a wild type (WT) recipient, WT donor and KO donor were labelled with CD8 alpha-Pacific Blue, TcR alpha/beta-FITC, Foxp3-PE, Ly5.1-PE-Cy5, CD4-PE-CY7, Thy1.2-APC and Ly5.2-APC-Cy7. This enabled the three sources of thymocytes to be analysed in a single tube.
Protocols
Immunophenotyping using directly conjugated monoclonal antibodies (mcabs)
Immunophenotyping with indirectly conjugated mcabs.
Immunophenotyping with a mixture of direct and indirectly conjugated mcabs.
Commonly used monoclonal antibody fluorophores are listed in fluorochromes used at ICMS table.
