Prevention of BoNT-B effects on primary afferent neurotransmitter release.

<p>Effective binding concentrations of BoNT-B and antibody were determined by titer assay on NativePAGE. Co-injection of BoNT-B and anti-BoNTB antibody was administered in mice 2 days prior to IPLT formalin, and spinal cord tissue was harvested at 10 minutes after formalin for analysis of NK1-R internalization. A) Immunoblot of BoNTB-antibody titer assay. BoNT-B was incubated with anti-BoNTB antibody (Ab) at dilutions 1∶1, 1∶5 and 1∶10. The effective binding concentrations were determined to be BoNTB 0.5 U/5 µL incubated with anti-BoNTB antibody at 1∶10 dilution, and BoNTB 1 U/5 µL incubated with antibody at 1∶5 dilution. B) Histogram of NK1-R internalization in the ipsilateral dorsal horn in animals that received IT BoNTB-antibody titer (n = 3), BoNTB 0.5 U–1 U (n = 3), and IT NaCl (n = 4). Bars represent mean ± SEM; P-value<0.05 significance.