Neuron-Specific Enolase

Chickens were immunized with two synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the NSE-2 gene product, but are shared between the human (NP_001966, NCBI) and rat (AAA41119, 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, and the concentrations of the eluates adjusted to 200 µ, g/ml. Finally, equal volumes of each of the two affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.

1:2000-1:5000 for immunohistochemistry and immunocytochemistry using 2% paraformaldehyde-fixed tissues or cells. 1:5000-1:10000 for western blots.