Baker-Herman, T.L., Fuller, D.D., Bavis, R.W., Zabka, A.G., Golder, F.J., Doperalski, N.J., Johnson, R.A., Watters, J.J., and Mitchell, G.S.
Notes: The NT-3 Emax® ImmunoAssay System was used to examine NT-3 levels in adult rat spinal cord following intrathecal injection of siRNAs. The C3-C5 sections of the spinal cord were dissected after treatment and homogenized in a cold extraction buffer (Tris-buffered saline [pH 8.0], 1% NP-40, 10% glycerol, 5mM sodium metavanadate, 10mM PMSF, 100μg/ml aprotinin and 10μg/ml leupeptin). The homogenates were acid treated, cleared by centrifugation, and then analyzed with the NT-3 Emax® ImmunoAssay System. Results were normalized by a protein assay. The researchers also used Promega’s recombinant human BDNF for phrenic activity studies on rats. In these experiments BDNF was dissolved in artificial cerebral spinal fluid containing 0.1%BSA. (3043)