Journal: The Journal of Neuroscience
Article Title: Altered Ion Channels in an Animal Model of Charcot-Marie-Tooth Disease Type IA
Figure Lengend Snippet: Kv3.1b is a component of the PNS nodes in Trembler-J nerves. These are images of unfixed teased sciatic nerve fibers from WT and Trembler-J ( TrJ ) mice, labeled as indicated. In WT mice, a few nodes are Kv3.1b positive (apposed arrowheads in A ), whereas
Article Snippet: Frozen sections and teased fibers were permeabilized by immersion in -20°C acetone for 10 min, blocked at room temperature for 1 h with 5% fish skin gelatin containing 0.1% Triton X-100 in PBS, and incubated overnight at 4°C with various combinations of primary antibodies: rabbit antisera against KCNQ2 [1:200 ( )], Kv3.1b (1:100; Alomone Labs, Jerusalem, Israel), Kv1.2 (1:100; Alomone Labs), Nav1.6 [1:100 ( )], Nav1.8 [1:100 ( )], Nav1.8 [1:500 ( )], ankyrin-G [1:100 ( ; )], contactin [1:100 ( )], ezrin-binding protein 50 kDa (EBP-50) (ab3452; 1:100; Abcam, Cambridge, MA), syndecan-3 [1:300 ( )], or Caspr [1:500 ( )]; and mouse monoclonal antibodies against panNav channels (K58/35; 1:250; Sigma, St. Louis, MO), Nav1.2 [1:100 ( ; )], Nav1.2 (1:50; Upstate Biotechnology, Lake Placid, NY), Caspr [1:50 ( )], annexin II light chain (1:50; BD Transduction Laboratories, San Jose, CA), or myelin-associated glycoprotein (MAG) (clone 513, 1:100; Boehringer Mannheim, Indianapolis, IN); rat monoclonal antibody against E-cadherin (1:50; Sigma); chicken antibody against βIV-spectrin [1:500 ( )]; and a soluble construct of receptor protein tyrosine phosphatase β conjugated to the human Fc fragment (RPTPβ-Fc) [1:3 ( )].
Techniques: Mouse Assay, Labeling