Neu-N (Fox 3)

Phosphate-buffered (10 mM) isotonic saline (0.9%, w.v) saline (PBS, pH 7.2) with 50% (v/v) glycerol and sodium azide (0.02%, w/v). The glycerol allows storage at -20C without freezing, the sodium azide is added as a preservative.

1:1000-1:2000 for immunohistochemistry and immunocytochemistry using 2% paraformaldehyde-fixed tissues or cells. Note that these dilutions are meant to serve as a starting point, and that optimal dilutions may vary.

Chickens were immunized with a synthetic peptide-keyhole limpet hemocyanin (KLH) conjugate. This synthetic peptide corresponded to the Fox-3 gene product (also known as the Neu-N antigen), but was shared between the human NP_001076044, NCBI) and mouse (NP_001020102, NCBI) sequences. After repeated injections into the hens, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, the concentrations of the eluate adjusted to 100 ug/ml, and the preparation filter-sterilized.