Guinea pig anti trpv1 antibody

Sections were rinsed in phosphate buffered saline (PBS) for 2 × 10 min, incubated in PBS containing 10% horse serum and 0.1% Triton X-100 for 30 min and processed for staining with the indirect immunofluorescence technique using the following antibodies: rabbit anti-InsR α subunit antibody (Santa Cruz Biotechnologies, Dallas, Tex., USA; 1:500), guinea pig anti-TRPV1 antibody (Neuromics, Edina, Minn., USA; 1:1500). The specificity of these antisera were assessed in DRG sections by the lack of staining with the TRPV1 antibody in specimens obtained from TRPV1 -/ -mice and by the failure of staining with the InsR antibody after preincubation with the immunising peptide supplied by the manufacturer (Baiou et al. 2007) . Donkey anti-rabbit IgG labelled with DL488 (1:500) and donkey anti-guinea pig IgG labelled with Cy3 (1:500) were used as secondary antibodies (all from Jackson Immunoresearch Laboratories, West Grove, Pa., USA). bWGA lectin was detected by using an extravidine-AMCA conjugate (Jackson Immunoresearch Laboratories). All antibodies were diluted in phosphate buffered saline containing Triton X100 (0.3%). Sections were incubated in the presence of the primary antibodies overnight followed by a 2h incubation with the secondary antibodies. Specimens were covered with Prolong Gold antifade mounting medium (Invitrogen, Carlsbad, CA, USA).