Table 1.

Antibodies used in this study.

Antigen	Immunogen	Host / type	Manufacturer	Specificity	Dilution
Calbindin	Rat recombinant Calbindin D28k	Rabbit polyclonal	Swant (Bellizona, Switzerland), cat. no. CB-38a	Detects single 28kDa band in WB of brain lysates from different species (mouse; rat; guinea pig; rabbit; Macaca fascicularis; zebrafish; chicken, Per manufacturer information). No staining in calbindin−/− mice (FJA lab: data not shown)	1:1,000- 1:5,000
VAChT	Synthetic peptide from rat VAChT (11 aa: 519–530)	Guinea pig polyclonal	Chemicon (Temecula, CA), cat. no. AB1588,	Detects a single band of 55–60 kDa band in WB of rat brain homogenates (Per manufacturer information). Co-localizes with ChAT in spinal cord sections (FJA lab: data not shown)	1:1,000
VAChT	Synthetic peptide from rat VAChT (20 aa: 511–530)	Goat polyclonal	Chemicon (Temecula, CA), cat. no. AB1578	Perfect colocalization with the immunoreactivity of AB1588 anti-VAChT (FJA lab: data not shown).	1:500
NeuN	Purified cell nuclei from mouse brain	Mouse monoclonal	Chemicon (Temecula, CA), cat. no. Mab377B	Labels specifically Fox-3 in tissue sections and a corresponding 46- 58 kDa band in WB of rat brain homogenates (Kim et al., 2009)	1:500–1:1,000
ChAT	Human placental enzyme	Goat polyclonal	Millipore cat. no. AB144P	Detects a single band of 68–70 kDa in WB of mouse brain lysates (Per manufacturer information).	1:250
Bassoon	Rat recombinant Bassoon produced using the sap7f cDNA clone (residues 738–1035)	Mouse monoclonal (mab7f)	University of Alabama hybridoma core. Obtained from Enzo Life Sci. cat. no. ADI- VAM-PS003.	Detects a single band of ~400 kDa band in WB of rat brain lysates (Per manufacturer information). No immunostaining in brains of animals lacking bassoon exons 4/5 (see text and Altrock et al., 2003).	1:4000

Abbreviations: NeuN, Neuronal Nuclear Protein; VAChT, Vesicular Acetylcholine Transporter; ChAT, choline acetyltransferase. WB, Western Blot (species and preparations tested in the WBs are fully described in Materials and Methods).