Protocol specific for LRRK2 Antibody (NB110-58771)

Immunostaining of frozen sections using streptavidin or Exvtravidin peroxidase and DAB

1. Make 10 micron frozen sections.
2. Fix tissue sections on slides in cold acetone, 10% buffered folrmalin or 4% paraformaldehyde for 8 to 10 min.
3. Rinse slides with PBS by immersion for 2 min. Repeat.
4. Block all slides with 5% milk in PBST for 30 min. at RT.
5. Remove blocking solution and add NB 110-58771 LRRK2 primary antibody 1:100 to 1:200 in PBS with 0.1% triton X-100 with 5% milk.
6. Incubate slides at 4degrees Celcius, overnight.
7. Rinse slides with PBST by immersion for 2 min. Repeat.
8. Add biotinylated donkey anti-rabbit secondary antibody (1:500 in PBST with 5% milk). Incubate 1 hour at RT.
9. Rinse with PBST by immersion 2 min. Repeat.
10. Add streptavidin-HRP or Extravidin-HRP. Incubate at RT for 1 hr.
11. Rinse with PBS by immersion for 1 min. Repeat.
12. Make DAB solution.
13. Add DAB to slides. Incubate 5-7 minutes.
14. Stop DAB reaction by immersion in water.
15. Counterstain sections, if desired.
16. Dehydrate,defat and coverslip slides.