3 Year Human Lung HOPX, AGER, and ACTA2 Confocal Imaging
Donor Tissue Kindly Provided by Dr. Gloria Pryhuber from the University of Rochester Medical Center
- For Frozen tissue, rinse 2X in PBS, then 5 min in 4% PFA/PBS, then rinse 1X in PBS.
- Briefly equilibrate slides in Antigen Retrieval Buffer.
- Antigen retrieval, pH 6.0 (times will vary according to microwave).
- 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).
- Microwave according to instructions on microwave.
- Cool on countertop, 15 min.
- Rinse with dH2O
- 1X PBS, 5 min.
- Block in 4% Donkey serum/PBS-T, 2 hours at RT.
- For rabbit anti- HOPX (SC-30216, Lot # C2212, Santa Cruz) dilute 1:200, For goat anti-AGER (AF1145, R&D Systems) dilute 1:100, for mouse anti-ACTA2 (