Alveolar Gas ExchangeArtboard 2@2x


Postnatal Day 14 C57BL6 HOPX, ABCA3, and ACTA2 Confocal Imaging



Immunofluorescence- HOPXABCA3, and ACTA2


Purpose:  Stain slides of 5µm paraffin sections of mouse lung C57BL6/J PND14 for HOPXABCA3, and ACTA2 with antigen retrieval.

Day 1

  1. For Frozen tissue, rinse 2X in PBS, then 5 min in 4% PFA/PBS, then rinse 1X in PBS.
  2. Briefly equilibrate slides in Antigen Retrieval Buffer.
  3. Antigen retrieval, pH 6.0 (times will vary according to microwave).
  4. 10 mM sodium citrate, pH 6.0, and heat in a microwave at 96oC. *We usually do a series of three runs (6-7 minutes each run) to equal the time/temp because of evaporation (refill coplin jars with dH2O).
  5. Microwave according to instructions on microwave.
  6. Cool on countertop, 15 min.
  7. Rinse with dH2O
  8. 1X PBS, 5 min.
  9. Block in 4% Goat serum/PBS-T, 2 hours at RT.
  10. For rabbit anti-HOPX (