Table 1.

Antibodies and DNA-binding dyes used

Target (against) or DNA-binding dyeComment on antibody or dye use/target/target functionCompanyCatalog #HostmAb/pAbFinal conc (μg/ml)
BrdUProliferation markerAbcam, Cambridge, UKab6326 [clone BU1/75 (ICR1)]RatmAb2 μg/ml
PCNAProliferation markerSanta Cruz Biotechnology, Santa Cruz, CAsc-56 (clone PC10)MousemAb2 μg/ml
Phospho-histone H3 (ser10)Proliferation markerMillipore, Bedford, MA06-570RabbitpAb5 μg/ml
Pax6Transcription factorCovance, Princeton, NJPRB-278PRabbitpAb1:400
p27Kip1Cell-cycle inhibitorBD Biosciences, San Jose, CA610242 (clone 57)MousemAb1.25 μg/ml
S100Calcium-binding protein expressed in various neural cell types; cytosolicDako, Carpinteria, CAZ0311RabbitpAb1:400
Type III β-tubulinNeuron-specific tubulinPromega, Madison, WIG712AMousemAb1.25 μg/ml
Hoechst 33342Binds double-stranded DNA; labels all nucleiSigmaB22611 μg/ml
DAPISame as Hoechst 33342SigmaD84171 μg/ml
Syto 24Same as Hoechst 33342Invitrogen, Carlsbad, CAS755950 nm
Mouse IgGSecondaryInvitrogenA11029Goat4 μg/ml
Mouse IgGSecondaryInvitrogenA11031Goat4 μg/ml
Rat IgGSecondaryInvitrogenA11077Goat2 μg/ml
Rabbit IgGSecondaryInvitrogenA11008Goat4 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.