Neuron-Specific Enolase (NSE)
Antibody Concentrations: 200 µg/ml. Two different affinity-purified anti-peptide antibodies were combined to make this product. The concentrations of each of the two antibodies was 100 µg/ml (based on Bradford assay readings using bovine serum albumin as a standard).
Volume: Regular vials contain 1000 µl of this antibody mixture; Sampler vials contain 200 ul of this mixture.
Buffer: Phosphate-buffered (10 mM) isotonic (0.9%, w/v) saline (PBS, pH 7.2) with sodium azide (0.02%, w/v) added as a preservative.
Quality Control: Each of the two antibodies were analyzed by immunohistochemistry (at a concentration of 3 ug/ml) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.
Recommended Dilutions: 1:2000-1:5000 for immunohistochemistry and immunocytochemistry using 2% paraformaldehyde-fixed tissues or cells. 1:5000-1:10000 for western blots.
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 two affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.