Simultaneous quantitation of equine cytokine mRNAs using a multi-probe ribonuclease protection assay.
- Additional Document Info
- View All
A rapid multi-probe ribonuclease protection assay (RPA) was developed to quantitate equine-specific cytokine mRNA levels in activated equine monocyte-derived macrophages (EMDM) and equine peripheral blood mononuclear cells (EPBMC). Eleven template plasmids specific to 10 equine cytokine genes and the beta-actin gene were generated from which radiolabeled anti-sense RNA probes were produced. The multi-probe RPA simultaneously quantitated mRNA levels of equine IL-1alpha, IL-1beta, IL-6, IL-8, IL-10, IL-12 p35, IL-12 p40, IFN-gamma, TGF-1beta and TNF-alpha in EPBMC and EMDM with coefficients of variation as low as 0.03-0.08 (3-8%) when normalized to beta-actin expression. This sensitive and rapid assay provides a valuable tool for studies of equine immune responses.
author list (cited authors)
Lim, W., Edwards, J. F., Boyd, N. K., Payne, S. L., & Ball, J. M.
complete list of authors
Lim, W-S||Edwards, JF||Boyd, NK||Payne, SL||Ball, JM