Flow Cytometric Analysis of Cell Signaling Proteins

Methods in Molecular Biology

Volumn 717, Issue , 2011, Pages 155-169

  Suni, Maria A ^a   Maino, Vernon C ^a  

^a BD BIOSCIENCES   (United States)

Author keywords

Cell signaling; Intracellular staining; Multiparameter flow cytometry; PBMC; Phospho specific antibodies; Phosphorylation; Whole blood

Indexed keywords

PHOSPHOPROTEIN; PHOSPHOSPECIFIC ANTIBODY;

ANIMAL; ARTICLE; BLOOD; CYTOLOGY; FLOW CYTOMETRY; HUMAN; METHODOLOGY; MONONUCLEAR CELL; SIGNAL TRANSDUCTION; STAINING;

ANIMALS; ANTIBODIES, PHOSPHO-SPECIFIC; FLOW CYTOMETRY; HUMANS; LEUKOCYTES, MONONUCLEAR; PHOSPHOPROTEINS; SIGNAL TRANSDUCTION; STAINING AND LABELING;

EID: 80052677854     PISSN: 10643745     EISSN: 19406029     Source Type: Book Series    
DOI: 10.1007/978-1-61779-024-9_9     Document Type: Chapter

References (18)

1. Cohen, P. (2000) The regulation of protein function by multisite phosphorylation–a 25 year update, Trends Biochem. Sci. 25, 596–601.
2. Krutzik, P. O., Irish, J. M., Nolan, G. P., and Perez, O. D. (2004) Analysis of protein phosphorylation and cellular signaling events by flow cytometry: techniques and clinical applications, Clin. Immunol. 110, 206–221.
3. Fleisher, T. A., Dorman, S. E., Anderson, J. A., Vail, M., Brown, M. R., and Holland, S. M. (1999) Detection of intracellular phosphorylated STAT-1 by flow cytometry, Clin. Immunol. 90, 425–430.
4. Montag, D. T., and Lotze, M. T. (2006) Successful simultaneous measurement of cell membrane and cytokine induced phosphorylation pathways [CIPP] in human peripheral blood mononuclear cells, J. Immunol. Methods 313, 48–60.
5. Krutzik, P. O., and Nolan, G. P. (2003) Intracellular phospho-protein staining techniques for flow cytometry: monitoring single cell signaling events, Cytometry A 55, 61–70.
6. Varker, K. A., Kondadasula, S. V., Go, M. R., Lesinski, G. B., Ghosh-Berkebile, R., Lehman, A., et al. (2006) Multiparametric flow cytometric analysis of signal transducer and activator of transcription 5 phosphorylation in immune cell subsets in vitro and following interleukin-2 immunotherapy, Clin. Cancer Res. 12, 5850–5858.
7. Irish, J. M., Hovland, R., Krutzik, P. O., Perez, O. D., Bruserud, O., Gjertsen, B. T., et al. (2004) Single cell profiling of potentiated phospho-protein networks in cancer cells, Cell 118, 217–228.
8. Montag, D. T., and Lotze, M. T. (2006) Rapid flow cytometric measurement of cytokine-induced phosphorylation pathways [CIPP] in human peripheral blood leukocytes, Clin. Immunol. 121, 215–226.
9. Haas, A., Weckbecker, G., and Welzenbach, K. (2008) Intracellular phospho-flow cytometry reveals novel insights into TCR proximal signaling events. A comparison with Western blot, Cytometry A 73, 799–807.
10. Perez, O. D., Mitchell, D., Campos, R., Gao, G. J., Li, L., and Nolan, G. P. (2005) Multiparameter analysis of intracellular phosphoepitopes in immunophenotyped cell populations by flow cytometry, Curr. Protoc. Cytom. Chapter 6, Unit 6.20.
11. Chow, S., Hedley, D., Grom, P., Magari, R., Jacobberger, J. W., and Shankey, T. V. (2005) Whole blood fixation and permeabilization protocol with red blood cell lysis for flow cytometry of intracellular phosphorylated epitopes in leukocyte subpopulations, Cytometry A 67, 4–17.
12. Desplat, V., Lagarde, V., Belloc, F., Chollet, C., Leguay, T., Pasquet, J. M., et al. (2004) Rapid detection of phosphotyrosine proteins by flow cytometric analysis in Bcr-Abl-positive cells, Cytometry A 62, 35–45.
13. Disis, M. L., dela Rosa, C., Goodell, V., Kuan, L. Y., Chang, J. C., Kuus-Reichel, K., et al. (2006) Maximizing the retention of antigen specific lymphocyte function after cryopreser-vation, J. Immunol. Methods 308, 13–18.
14. Maecker, H. T., and Trotter, J. (2006) Flow cytometry controls, instrument setup, and the determination of positivity, Cytometry A 69, 1037–1042.
15. Maino, V. C., and Picker, L. J. (1998) Identification of functional subsets by flow cytometry: intracellular detection of cytokine expression, Cytometry 34, 207–215.
16. Maecker, H. T. (2004) Cytokine flow cytometry, Methods Mol. Biol. 263, 95–108.
17. Chow, S., Patel, H., and Hedley, D. W. (2001) Measurement of MAP kinase activation by flow cytometry using phospho-specific antibodies to MEK and ERK: potential for pharmacodynamic monitoring of signal transduction inhibitors, Cytometry 46, 72–78.
18. Tong, F. K., Chow, S., and Hedley, D. (2006) Pharmacodynamic monitoring of BAY 43-9006 (Sorafenib) in phase I clinical trials involving solid tumor and AML/MDS patients, using flow cytometry to monitor activation of the ERK pathway in peripheral blood cells, Cytometry B Clin. Cytom. 70, 107–114.