Anti-P-Zero Myelin Protein (PZO) Antibody

Human Myelin Protein Zero (Pz0) is a 28,400 dalton protein (258 amino acids) found in myelin of the PNS, but not the CNS. Expressed in myelinating Schwann cells, Po serves as a homophilic adhesion molecule, allowing adjacent laminae of the myelin sheath to undergo compaction. Po is also a marker of a subpopulation of migrating neural crest cells. Mutations of the MPZ gene product in humans is responsible for Charcot-Marie-Tooth type 1B neuropathies (HMSN1B) .

Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least twelve months, provided they remain sterile. For longer term storage, aliquot and freeze to avoid freeze-thaw of the antibody.

Chickens were immunized with two synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the Po gene product, but are shared between the human (NP_000521, NCBI) and mouse (NP_032649, 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 μg/mL. Finally, equal volumes of both of these affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.

Both of these anti-peptide antibodies were analyzed by immunohisto-chemistry (at a dilution of 1:2000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.