Immunization with alpha-synuclein peptide

Prepare immunization emulsions:

Prepare 200µg of M. tuberculosis H37Ra (BD Difco Adjuvant, Fisher Scientific, Cat #DF0638-60-7; Waltham, MA) in 1x PBS or in 100µg a-syn32-46 peptide (A&A peptides; San Diego, CA) dissolved in 1x PBS.

Immunize 8- to 12-week-old male and female mice subcutaneously with 50µL of emulsion into the chest area of the mouse, near the axillary lymph node at both front limbs for a totally of 100µL of emulsion injected per mouse.

The day of immunization is designated as day 0.

At day 14, boost each mouse with the same immunization dose at the same site.

Administer intravenous injections of 400ng of B. pertussis toxin (List Biological Laboratories, Cat #181; Campbell, CA) at 14- and 16-days post-initial immunization (day 0 and 2 post boost).

Monitor mice daily for their weight, food and fluid intake and neurological deficits and their weights are recorded daily. Mice are classified as sick if they displayed 12% reduction from 0 DPI weight and/or hunched posture with ruffled and ungroomed fur.