Accurate and reproducible measurements of RhoA activation in small samples of primary cells |
| |
| Authors: | Lylia Nini |
| |
| Institution: | a Department of Physiology and Pharmacology, University of Western Ontario, London, Ontario, Canada N6A 5C1
b Department of Pediatrics, Child Health Research Institute and Lawson Health Research Institute, University of Western Ontario, London, Ontario, Canada N6A 5C1 |
| |
| Abstract: | Rho GTPase activation is essential in a wide variety of cellular processes. Measurement of Rho GTPase activation is difficult with limited material, such as tissues or primary cells that exhibit stringent culture requirements for growth and survival. We defined parameters to accurately and reproducibly measure RhoA activation (i.e., RhoA-GTP) in cultured primary keratinocytes in response to serum and growth factor stimulation using enzyme-linked immunosorbent assay (ELISA)-based G-LISA assays. We also established conditions that minimize RhoA-GTP in unstimulated cells without affecting viability, allowing accurate measurements of RhoA activation on stimulation or induction of exogenous GTPase expression. |
| |
| Keywords: | RhoA Primary cell Keratinocyte |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
