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How do you stain paraffin-embedded tissue with a phalloidin"conjugate?

Posted August 30, 2018


Answer

Reagents

Fixative - 3.7% paraformaldehyde in PBS
Permeabilization Buffer – PBS+ 0.2% Triton X-100
Blocking Buffer – PBS+1% BSA+0.05% Triton X-100

Procedure:

  1. Dewax the tissue.
  2. Add Permeabilization Buffer for one or two hours, slowly rotate the tissue sections .
  3. After Permeabilization, wash the tissue sections with three changes of PBS+ 0.2% Triton X-100 Buffer, 5-10 min/wash.
  4. Add Blocking Buffer to block the nonspecific fluorophore binding sites for 1-2 hours.
  5. 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.
  6. Wash with excess PBS+ 0.2% Triton X-100.
  7. Observe under microscope.
Additional resources

Phalloidin-iFluor™ 488 Conjugate