Chicken anti nf200

Antibodies used were rabbit anti‐S100 (Dako, Carpenteria, CA, USA) cat. no. Z0311, RRID:AB_10013383) diluted 1:1000; chicken anti‐NF200 (Millipore (Billerica, MA, USA) cat. no. AB5539, RRID:AB_177520 diluted 1:1000; anti‐rabbit Alexa Fluor 488 (Thermo Fisher Scientific, Waltham, MA, USA, cat. no. A‐11008, RRID:AB_143165) diluted 1:800; and anti‐chicken Alexa Fluor 647 (Thermo Fisher Scientific, Waltham, MA, USA, cat. no. A‐21449, RRID:AB_2535866) diluted 1:800).

Human and mouse DRG sections of 20 µm thickness were washed with PBS, fixed with 4% PFA for 30 min and blocked with 10% goat serum/0.1% TritonX-100 in PBS for 1 h at room temperature.
Primary antibodies (mouse ant-Tuj1 from Covance, 1:750; chicken anti-NF200 from Millipore, 1:25,000; mouse anti-NF200 (clone N52) from Sigma, 1:600 and rabbit anti-Brn-3a from Merck, 1:200) were diluted in 3% goat serum/0.1% TritonX-100 in PBS and applied over night at 4 °C. After rinsing 3 times with 0.1%Triton/PBS, sections were incubated with secondary antibodies (Alexa dyes 488 and 555 from dianova) and DAPI (diluted in 3% goat serum/0.1% TritonX-100) for 3 h at room temperature. Sections were washed with 0.1% TritonX-100/PBS and PBS and mounted with ImmuMount.