Neurofilament, Light Chaiin (NF-L)

Phosphate-buffered (10 mM) isotonic (0.9%, w/v) saline (PBS, pH 7.2) with sodium azide (0.02%, w/v) added as a preservative.

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

Chickens were immunized with three synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the NF-L gene product, but are shared between the human (P07196, NCBI) and mouse (P08551, NCBI) protein sequences. After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, and the concentrations of the eluates adjusted to 300 µ, g/ml. Finally, equal volumes of each of the three affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.