Anti-Nestin Antibody

Mouse Nestin is a 206,994 dalton protein (1864 amino acids) expressed in neural crest cells, CNS neural stems cells, as well as a few non-neural cell types in the embryo, including cells within the pancreatic islets of Langerhans and the limb bud. Nestin is an intermediate filament-associated protein, and contributes to the cytoskeleton.

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 three synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the mouse nestin gene product (NM_NP_057910), but are shared with the rat (NM_012987, NCBI) protein sequence. 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 μg/mL. Finally, equal volumes of each of the three affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.

Each of the three antibodies were analyzed by immunohistochemistry (at a dilution of 1:20000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.