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Updated: May 13, 2026
Flow Cytometry Protocols for Surface and Intracellular Antigen Analyses of Neural Cell Types
Published on: December 18, 2014
Optimized flow cytometry protocol for analysis of surface expression of interleukin-1 receptor types I and II
Filipp Filippovich Vasilyev1, Julia Anatolievna Lopatnikova, Sergey Vitalievich Sennikov
1Laboratory of Molecular Immunology, Federal State Budgetary Institution "Research Institute of Clinical Immunology", Russian Academy of Medical Sciences Siberian Branch, 14, Yadrincevskaja str., 630099, Novosibirsk, Russia, vasilyevmd@gmail.com.
Abstract:
The biological effects of interleukin (IL)-1 are realized through binding to specific membrane-bound receptors. The efficiency of IL-1 action depends on the number of receptors on the cell. We determined the percentage of cells that express IL-1 receptor type I (IL-1RI) and IL-1 receptor type II (IL-1RII) by flow cytometry using phycoerythrin (PE)-labelled antibodies to the IL-1Rs, and the mean absolute number of membrane-bound IL-1Rs per cell using QuantiBRITE PE calibration beads. We showed that different subpopulations of immunocompetent cells expressed different numbers of molecules of membrane-bound IL-1RI and IL-1RII. We also established that when cells were stimulated with bacterial lipopolysaccharide, there was a significant increase in the number of IL-1RI expressed, and a significant decrease in the mean number of IL-1RII molecules per cell. Determination of the mean number of membrane-bound IL-1R molecules using this protocol enables us to obtain precise and reproducible data that are necessary for full evaluation of expression levels.
