UCSF

IF paraffin slides

IF Staining with Paraffin Section Slide

Procedure

  1. Heat slides in an oven at 60°C for 45 minutes. (usually do a day before)
  2. Deparaffinize/hydrate using the following series of washes: Xylenes 3X 5 min 100% EtOH 2X 2 min 95% EtOH 2X 2 min 80% EtOH 1X 2 min 70% EtOH 1X 2 min dH2O 2X 3 min + shaker PBS 1X 5 min + shaker
  3. Microwave slide in boiling 10mM Sodium Citrate Buffer. Fill chamber to brim with buffer. Microwave 10 minutes on high power in buffer and allow 20 minutes to cool down at RT.
  4. Fix with 200ul of 4% paraformaldehyde/PBS for 20 min at RT (fix time)
  5. Wash with PBS
  6. Permeabilize with 0.1% Triton x-100/PBS for 3 min at RT (fix time)
  7. Wash with PBS
  8. Treated with goat serum/PBS (1:200) for 30 min at RT.
  9. Wash with PBS
  10. Added primary Antibody in goat serum/PBS for 1 hr at RT
  11. Wash with PBS
  12. Added goat anti-rabbit 594 or anti-mouse for 30 min at RT.
  13. Wash with PBS
  14. Mounting the slide with reagent contain DAPI.
  15. Store at dark box at 4ºC. Good for one week.