| | |  | | | | Record from Web of Science® | | | | | | |
| Simultaneous measurement of multiple active kinase states using polychromatic flow cytometry |
|
|
| Author(s): Perez OD, Nolan GP |
| Source: NATURE BIOTECHNOLOGY Volume: 20 Issue: 2 Pages: 155-162 Published: FEB 2002 |
| Times Cited: 119 References: 23 |
| Abstract: Intracellular assays of signaling systems have been limited by an inability to correlate functional subsets of cells in complex populations on the basis of active kinase states. Such correlations could be important in distinguishing changes in signaling status that arise in rare cell subsets during functional activation or in disease manifestation. Here we demonstrate the ability to simultaneously detect activated kinase members of the mitogen-activated protein kinases family (p38 MAPK, p44/42 MAPK, JNK/SAPK), members of cell survival pathways (AKT/PKB), and members of T-cell activation pathways (TYK2), among others, in subpopulations of complex cell populations by multiparameter flow-cytometric analysis. We demonstrate the utility of these probes in identifying distinct signaling cascades for (1) both artificial and physiological stimulatory conditions of peripheral blood mononuclear cells (PBMCs), (2) cytokine stimulation in human memory and naive lymphocyte subsets as identified by five differentiation markers, and (3) ordering of kinase activation in potential signaling hierarchies. Polychromatic flow-cytometric active kinase measurements demonstrate that multidimensional analysis of signaling pathways can provide functional signaling pathway assessment on a single-cell level and allow for potential correlation with biological and clinical parameters. |
| Document Type: Article |
| Language: English |
| Reprint Address: Nolan, GP (reprint author), Stanford Univ, Sch Med, Dept Microbiol & Immunol, Stanford, CA 94305 USA |
Addresses:
1. Stanford Univ, Sch Med, Dept Microbiol & Immunol, Stanford, CA 94305 USA
2. Stanford Univ, Sch Med, Baxter Lab Genet Pharmacol, Stanford, CA 94305 USA
3. Stanford Univ, Sch Med, Dept Mol Pharmacol, Stanford, CA 94305 USA |
| Publisher: NATURE AMERICA INC, 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA |
| Subject Category: Biotechnology & Applied Microbiology |
| IDS Number: 517FT |
| ISSN: 1087-0156 |
|
| | | | | | | | | Record from Web of Science® | | | | | | | | | |