Table 1.
Antibodies and DNA-binding dyes used
| Target (against) or DNA-binding dye | Comment on antibody or dye use/target/target function | Company | Catalog # | Host | mAb/pAb | Final conc (μg/ml) |
|---|---|---|---|---|---|---|
| BrdU | Proliferation marker | Abcam, Cambridge, UK | ab6326 [clone BU1/75 (ICR1)] | Rat | mAb | 2 μg/ml |
| PCNA | Proliferation marker | Santa Cruz Biotechnology, Santa Cruz, CA | sc-56 (clone PC10) | Mouse | mAb | 2 μg/ml |
| Phospho-histone H3 (ser10) | Proliferation marker | Millipore, Bedford, MA | 06-570 | Rabbit | pAb | 5 μg/ml |
| Pax6 | Transcription factor | Covance, Princeton, NJ | PRB-278P | Rabbit | pAb | 1:400 |
| p27Kip1 | Cell-cycle inhibitor | BD Biosciences, San Jose, CA | 610242 (clone 57) | Mouse | mAb | 1.25 μg/ml |
| S100 | Calcium-binding protein expressed in various neural cell types; cytosolic | Dako, Carpinteria, CA | Z0311 | Rabbit | pAb | 1:400 |
| Type III β-tubulin | Neuron-specific tubulin | Promega, Madison, WI | G712A | Mouse | mAb | 1.25 μg/ml |
| Hoechst 33342 | Binds double-stranded DNA; labels all nuclei | Sigma | B2261 | 1 μg/ml | ||
| DAPI | Same as Hoechst 33342 | Sigma | D8417 | 1 μg/ml | ||
| Syto 24 | Same as Hoechst 33342 | Invitrogen, Carlsbad, CA | S7559 | 50 nm | ||
| Mouse IgG | Secondary | Invitrogen | A11029 | Goat | 4 μg/ml | |
| Mouse IgG | Secondary | Invitrogen | A11031 | Goat | 4 μg/ml | |
| Rat IgG | Secondary | Invitrogen | A11077 | Goat | 2 μg/ml | |
| Rabbit IgG | Secondary | Invitrogen | A11008 | Goat | 4 μg/ml |
Primary and secondary antibodies and DNA-binding dyes used in this study are listed. ″Final conc″ is the concentration of the antibody in the solution in which the tissue was incubated. For antibodies for which the vendor stated the stock concentration, final concentration is given in micrograms per milliliter. When the stock concentration was unknown, the dilution is stated. In one experiment (shown in Fig. 1F), the BrdU antibody was used at a final concentration of 0.5 μg/ml; with this single exception, antibodies were used at the concentration shown in the Table. Note that although BrdU is listed in the table as a ″Proliferation Marker″ (and indeed its most common use is in studying proliferative characteristics of cell populations), DNA synthesis (and hence BrdU labeling) can occur in the absence of cell division [Nowakowski and Hayes (2000) and references therein]. mAb, Monoclonal antibody; pAb, polyclonal antibody; PCNA, proliferating cell nuclear antigen.