Whole blood processing for measurement of signaling proteins by flow cytometry.
Signal transduction pathways link external stimuli with cellular responses, which normally regulate cell proliferation, death, and differentiation. The study of signal transduction was revolutionized through the development of phospho-specific antibodies that recognize proteins only when they are phosphorylated at specific sites. As discussed by Nolan and co-workers (UNIT ), one of the unique features of flow cytometry is its ability to perform correlated measurements of multiple phosphorylation states at the single cell level. This provides insight into the complexity of signaling networks that is not obtained by standard biochemical techniques. Furthermore, in combination with other phenotypic markers, flow cytometry can measure alterations in signaling pathways in subpopulations of cells. This clearly has wide potential for studying disorders of the hematopoietic and immune systems.
Chow
Sue
S
Division of Applied Molecular Oncology, Ontario Cancer Institute, Princess Margaret Hospital, Toronto, Ontario, Canada.
Hedley
David
D
Shankey
T Vincent
TV
D015703:Antigens, CD / Q000097:blood; D001774:Blood Chemical Analysis / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000097:blood*; D015470:Leukemia, Myeloid, Acute / Q000097:blood; D020935:MAP Kinase Signaling System; D051058:Oncogene Protein v-akt / Q000097:blood; D019869:Phosphatidylinositol 3-Kinases / Q000097:blood; D015398:Signal Transduction
D015703:Antigens, CD; D047908:Intracellular Signaling Peptides and Proteins; D019869:Phosphatidylinositol 3-Kinases; D051058:Oncogene Protein v-akt