Two antipeptide antibodies were generated in chickens against a sequence shared between the rat (AAA41119) and human (NP_001966) gene products. Antibodies were affinity-purified and mixed in equal concentrations. These antibodies have been validated with human, mouse and rat tissues.
Immunocytochemical staining of NSE (Aves Labs, GREEN, 1:1000 dilution) and Glial Fibrillary Acidic Protein (GFAP, rabbit, EnCor, 1:1000 dilution). Photomicrograph by Dr. Gerry Shaw (EnCor Biotechnology).
Anti-Neuron-Specific Enolase Antibody
Neuron-Specific Enolase (NSE) Type 2
Human NSE-2 (EC 18.104.22.168) is a 47,138 dalton protein (434 amino acids) expressed in neurons of the peripheral nervous system (PNS) and central nervous system (CNS). NSE-2 catalyzes the conversion of 2-phospho-D-glycerate into phosphoenol pyruvate and is an essential enzyme in energy metabolism in nervous tissues.
ICC, IHC, WB
Human, Mouse, Rat
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 two synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the NSE-2 gene product, but are shared between the human (NP_001966, NCBI) and rat (AAA41119, NCBI) sequences. After repeated injections into the hens, 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 each of the three 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 1:5000) 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.