Three different antipeptide antibodies were generated in chickens against sequences shared between the mouse (NP_035831) and human (NP_003371) gene products. Antibodies were affinity-purified and mixed in equal concentrations. These antibodies have been validated with human, mouse and rat tissues.
Immunostaining of adult mouse cerebellum, showing staining in the granule cell layer and white matter tracts. Anti-vimentin antibody was used at a dilution of 1:1000; HRP-labeled goat anti-chicken IgY was used at a dilution of 1:500.
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.
ICC, IHC, WB
Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.
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 filtersterilized.
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.
These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.
UniProt (Immunogen Species): P08670
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