GAD (Glutamic Acid Decarboxylase-67)
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: The antibody was analyzed by immunohistochemistry (at a dilution of 1:1000) 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 a synthetic peptide/keyhole limpet hemocyanin (KLH) conjugate corresponding to a region of the Glutamic Acid Decarboxylase (GAD-67) EC 4.1.1.15) shared between the mouse (P48318), rat (NP_058703) and human (Q99259) gene products. After repeated injections into the hens, immune eggs were collected, and the IgY fraction were purified from the yolks. These IgY fractions were then affinity-purified using peptide columns, the concentration of the eluate adjusted to 100 µg/ml, and the preparation was filter-sterilized.