How do you stain paraffin-embedded tissue with a phalloidin"conjugate?
Posted August 30, 2018
Fixative - 3.7% paraformaldehyde in PBS
Permeabilization Buffer – PBS+ 0.2% Triton X-100
Blocking Buffer – PBS+1% BSA+0.05% Triton X-100
- Dewax the tissue.
- Add Permeabilization Buffer for one or two hours, slowly rotate the tissue sections .
- After Permeabilization, wash the tissue sections with three changes of PBS+ 0.2% Triton X-100 Buffer, 5-10 min/wash.
- Add Blocking Buffer to block the nonspecific fluorophore binding sites for 1-2 hours.
- After blocking, place the slides in a humidity chamber and add 1X Phalloidin–Conjugate in PBS Buffer + 1% BSA to the surface of the tissue section. Incubate the covered slides in the humidity chamber for one or two hours at RT or 37oC, avoid light.
- Wash with excess PBS+ 0.2% Triton X-100.
- Observe under microscope.