TABLE 1.
Primary Antibodies
| Antiserum | Immunogen | Source (cat. no.) | Working dilution | Specificity |
|---|---|---|---|---|
| Goat polyclonal anti-doublecortin (DCX) | Synthetic peptide corresponding to AA 385–402 at the c-terminus of human doublecortin | Santa Cruz Biotechnology, Santa Cruz, CA (SC-8066) | 1:500 | A single band at 40 kDa was detected on a Western blot of adult rat olfactory bulb.(Brown et al., 2003). |
| Rabbit polyclonal anti-growth associated protein 43 (GAP-43) | Synthetic peptide corresponding to AA 216–226 of rat/mouse GAP-43. | Novus Biologicals, Littleton, CO (NB300-143) | 1:1,000 | A band at 47 kDa is detected in Western blots of brain (manufacturer’s technical information). Staining with GAP-43 was consistent with the staining pattern previously described, staining the lower part of the OE (Iwema and Schwob, 2003; Rodriguez-Gil and Greer, 2008). |
| Goat polyclonal anti-olfactory marker protein (OMP) | Purified natural Rat OMP | Wako, Dallas, TX (#544-10001) | 1:2,000 | This antibody recognized a single band at 19 kDa on a Western blot of mouse brain. Preabsorption blocks all staining (Baker et al., 1989; Koo et al., 2004). |
| Rabbit polyclonal anti-brain lipid binding protein | Recombinant human full length BLBP protein | Millipore, Billerica, MA (AB9558) | 1:2,000 | A single band at 14.5 kDa on a Western blot of mouse cerebellum and cortex (Feng et al., 1994). |
| Rabbit polyclonal anti-peripherin | Electrophoretically pure trp-E-peripherin fusion protein containing all but the four N-terminal amino acids of rat peripherin | Millipore, Billerica, MA (AB1530) | 1:2,000 | This antibody recognizes a single band at 57 kDa on a Western blot from PC12 lysates (manufacturer’s technical information) |
| Mouse monoclonal anti-β-tubulin III, clone SDL.3D10 | Synthetic peptide corresponding to the C-terminal sequence of human b-tubulin isotype III coupled to BSA. The exact sequence is: C-E S E S Q G P K. | Sigma, St. Louis, MO (T8660) | 1:250 | A single 46 kDa band was detected on a Western blot from bovine brain; no cross-reactivity with other tubulin isotopes (manufacturer’s technical information; Banerjee et al., 1990). |
| Rabbit polyclonal anti-gonadotrophin releasing hormone (GnRH) | Synthetic peptide corresponding to residues pyroE(22) H W S Y G L R P G- NH2 (32) of mouse GnRH | Affinity BioReagents (now Thermo Scientific Pierce Antibodies, Rockford, IL) (PA1-121) | 1:1,000 | Western blot from mouse brain has an expected band at 10.4 kDa. Staining with GnRH produced the expected pattern of staining in the developing embryos (Leupen et al., 2003; Schwanzel-Fukuda and Pfaff, 1989; Wray et al., 1989) |
| Goat polyclonal anti-Delta/Notch-like EGF-related Receptor (DNER) | Purified, NSO-derived, recombinant mouse DNER extracellular domain; AA 26–640. | R&D Systems, Minneapolis, MN (AF2254) | 1:200 | Western blot from rat brain shows a 78 kDa band (manufacturer’s technical information). Staining pattern showed labeling of developing neurons (confirmed with double staining using established neuronal markers. Cells were adjacent to glial cells consistent with the expected role for this molecule in glial cell differentiation (Eiraku et al., 2005). |
| Goat polyclonal anti-LIM homeobox do- main 2 (Lhx2) | Peptide mapping at the C-terminus of Lhx2 of human origin. Specific sequence: VTSVLTSVPGNLEGHEPHSP | Santa Cruz Biotechnology, Santa Cruz, CA (SC-19344) | 1:1,000 | The staining pattern seen was consistent with the expected, published staining pattern in the OE and MM cells, previously described as pioneer neurons (Ikeda et al., 2007). Western blots of mouse brain have an apparent band at 43 kDa. |