43 Year-Old Human Lung NKX2.1, AGER, and ACTA2 Confocal Imaging
Gene |
|||||
Nkx2-1 | LungGens | Ensemble | Genecards | NCBI | The Human Protein Atlas |
AGER | LungGens | Ensemble | Genecards | NCBI | The Human Protein Atlas |
Acta2 | LungGens | Ensemble | Genecards | NCBI | The Human Protein Atlas |
Donor Tissue Kindly Provided by Dr. Gloria Pryhuber from the University of Rochester Medical Center
Immunofluorescence- NKX2.1, AGER, and ACTA2
Purpose: Stain slides of 7µm frozen sections of 43-year-old human lung, courtesy of Dr. Scott Randell, University of North Carolina, for NKX2.1, AGER, and ACTA2 with antigen retrieval.
Day 1
- 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- NKX2.1 (RB TTF-1 1231, Lot# 0912A) dilute 1:1000, For rat anti-