Goat anti chicken igg conjugated alexa fluor 594

To generate Kcns1 antibody, the C-terminal end region of mouse Kcns1 (aa 469-497, NP_032461.2) was PCR amplified and subcloned into pGEX-4T vector (glutathione S-transferase fusion protein, Amersham Pharmacia Biotech). Glutathione S-transferase fusion proteins were purified using glutathione-Sepharose 4B resin (Amersham Pharmacia Biotech) and used to immunize rabbit. Kcns1 antibody was affinity purified using glutathione S-transferase fusion protein. Primary antibodies and their dilutions used in this study were: rabbit anti-Kcns1 (1:2000), mouse anti-NeuN (1:1000, Abcam #ab104224), chicken anti-β3tubulin (1:1000, Abcam #ab41489), chicken anti-peripherin (1:500, Abcam #ab39374), goat anti-CGRP (1:1000, Abcam #ab36001), chicken anti-NF200 (1:1000, Merck Millipore AB5539), mouse anti-Na_v1.8 (1:1000, Neuromab N134/12), goat anti-CSF1 (1:1000, R&D Systems #AF416), and rabbit anti-ATF3 (1:500, Santa Cruz Biotechnology #sc-188). Secondary antibodies used were donkey anti-mouse IgG-conjugated Alexa Fluor 488 or 594, goat anti-chicken IgG-conjugated Alexa Fluor 594, donkey anti-rabbit IgG-conjugated Alexa Fluor 488 or 594, and donkey anti-goat–conjugated Alexa Fluor 594 (all 1:1000, Invitrogen). IB4 binding was visualized using biotin-conjugated IB4 (1:200, Sigma L2140) and Avidin-FITC (1:500, Sigma).