NF-L (Neurofilament Protein, 68 kDa)
Antibody Concentrations: 300 µg/ml.
Volume: Regular vials contain 1000 µl of this antibody mixture; Sampler vials contain 200 µl 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: 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.
Recommended Dilutions: 1:1000-1:2000 for immunohistochemistry and immunocytochemistry using 2% paraformaldehyde-fixed tissues or cells. Please note that these dilutions are meant to serve as starting points, and that optimal dilutions may vary.
Chickens were immunized with three synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the NF-L gene product, but are shared between the human (P07196, NCBI) and mouse (P08551, 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.