Anti-Vimentin Protein Antibody

Human Vimentin is a 53,521 dalton protein (466 amino acids) expressed in many cell types of mesodermal origin and is one of the intermediate filament types that form the cytoskeleton. In the embryonic nervous system, however, Vimentin is expressed by migrating neural crest cells forming the PNS and by neural stem cells of the CNS. In some forms of glioblastoma, Vimenin is re-expressed by the tumor cells and serves as a general indicator of their undifferentiated state.

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 vimentin gene product, but are shared between the human (NP_003371, NCBI) and mouse (NP_035831, 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 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 1:2000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.