Table 1.
Primary antibodies used. LCM 29 and LCM3 are mAbs that recognize all lamprey glial keratin and all lamprey neurofilament subunits, respectively (see Methods). They have been characterized previously with regard to specificity in lamprey CNS (Merrick et al., 1995; Pleasure et al., 1989). The biotin-conjugated anti-HuD antibody was purified from the serum of a patient with paraneoplastic encephalomyelitis, and was kindly provided by Dr. Joseph Dalmau, Department of Neurology, University of Pennsylvania, now at University of Barcelona Hospital Clinic.
| Antigen | Immunogen | Host and Type | Manufacturer | Dilution |
|---|---|---|---|---|
| Bromodeoxyuridine | Bromodeoxyuridine-bovine serum albumin |
Mouse monoclonal |
Chemicon (Temecula, CA), MAB3424 |
1:200 |
| Bromodeoxyuridine | Bromodeoxyuridine coupled to keyhole limpet hemocyanin |
Sheep polyclonal |
Abcam, ab1893 | 5 μg/ml |
| Neuronal Protein HuC/HuD and Hel- N1 |
Human neuronal protein | Mouse monoclonal |
Molecular Probes, 16A11 |
1:100 |
| Cytokeratin | Lamprey cytoskeleton | Mouse monoclonal | Selzer lab, LCM29 | 1:100 |
| Neuronal protein HuD |
Recombinant Human HuD protein |
Human polyclonal | Dr. Josep Dalmau | 1:500 |
| Neurofilaments | Lamprey cytoskeleton | Mouse monoclonal | Selzer lab, LCM 3 | 1:100 |
| Acetylated α - tubulins |
Outer arm of sea urchin | Mouse monoclonal |
Sigma-Aldrich, 6-11B-1 |
1:500 |