Microtubule-Associated Protein (MAP-2)

Chickens were immunized with two synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the MAP-2 gene product, but are shared between the human (NP_002365, NCBI) and mouse (P20357, NCBI) 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 200 ug/ml. Finally, equal volumes of both of these affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.

1:1000-1:2000 for immunohistochemistry and immunocytochemistry using 2% paraformaldehyde-fixed tissues or cells. 1:2000-1:5000 for western blots. Note that these dilutions are meant to serve as starting points, and that optimal dilutions may vary.