Netrin
Antibody Concentrations: 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 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: This antibody preparation was analyzed by immunohistochemistry (at a concentration of 3 µg/ml) 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. Note that these dilutions are meant to serve as starting points, and that optimal dilutions may vary.
Chickens were immunized with a synthetic peptide-keyhole limpet hemocyanin (KLH) conjugate. This synthetic peptide corresponded to the netrin-1 gene product, but was shared between the human (O_095631, NCBI) and mouse (O_09118, 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, the concentrations of the eluate adjusted to 100 µg/ml, and the preparation filter-sterilized.