Other Reagents to Consider:
Chien-Cheng Liu, Yong-Jing Gao, Hao Luo, Temugin Berta, Zhen-Zhong Xu, Ru-Rong Ji & Ping-Heng Tan. Interferon alpha inhibits spinal cord synaptic and nociceptive transmission via neuronal-glial interactions. Scientific Reports 6, Article number: 34356 (2016). doi:10.1038/srep34356
Animals were terminally anesthetized with isoflurane and perfused through the ascending aorta with saline followed by 4% paraformaldehyde with 1.5% picric acid in 0.16 M PB. After the perfusion the L4-L5 spinal cord segments and DRGs were collected and postfixed in the same fixative overnight. Spinal cord sections (30 μm, free-floating sections) and DRG sections (20 μm) were cut in a cryostat and processed for immunofluorescence. Tissue sections were blocked with 2% goat serum, and incubated over night at 4 °C with the following primary antibodies: IFN-α/βRβ (rabbit, 1:500, Santa Cruz, sc-30015, M-300), IFN-α antibody (rabbit, R&D, 1:1000), GFAP antibody (mouse, 1:5000, Chemicon), OX-42 antibody (mouse, 1:5000, Serotec), NeuN antibody (mouse, 1:5000, Chemicon), CGRP antibody (chicken, 1:2000, Neuromics), substance P antibody (guinea pig, 1:500, Neuromics), NF-200 antibody (mouse, 1:2000, Sigma), NK-1 antibody (rabbit, 1:2000; Neuromics) and phosphorylated ERK antibody (pERK1/2, rabbit, Cell Signaling, 1:500), as well as FITC-conjugated IB4 (1:200, Sigma). The sections were then incubated for 1 h at room temperature with Cy3- or FITC- conjugated secondary antibodies (1:300, Jackson immunolab). For double immunofluorescence, sections were incubated with a mixture of polyclonal and monoclonal primary antibodies followed by a mixture of FITC- and CY3-congugated secondary antibodies...
Shigekazu Sugino, Mohamed Farrag, Victor Ruiz-Velasco. Gα14 subunit-mediated inhibtion of voltage-gated Ca2+ and K+ channels via neurokinin-1 receptor in celiac-superior mesenteric ganglion neurons. Journal of Neurophysiology Published 3 February 2016 Vol. no. , DOI: 10.1152/jn.00980.2015
...172 The neurons were incubated with the rabbit anti-NK-1 receptor (1:100; Neuromics, Edina,MN) 173 ... No. 195 RA-25001, Neuromics) ...
- Neurokinin-1 (NK 1) Receptor-Detects a band at 80-90 kDa on Western blots of
membranes prepared from cells transfected with the rat substance P receptor (Vigna et al., 1994); stainingin rat spinal cord was blocked by preabsorbing the antiserum with the immunizing peptide (Mantyh et al., 1995)-Dilution 1:2,000
Substance P-The distribution of immunoreactivity in rat spinal cord is identical to that described previously (Hunt et al.,1981); in dual-labeling experiments, it labels the same structures as a polyclonal rabbit anti-SP (1:1,000; Peninsula/Bachem; T-4107; data not shown)