FACS analysis for E15.5 mouse kidneys (Version 1.0)

Version

1.0

Notice

This page is the corresponding protocol tomestone page generated as part of the ATLAS-D2K shutdown in July 2025. Many links on this page may be broken.

Authors

Steve Potter

Keywords

[‘FACS’]

Subjects

[‘Isolation, purification and separation’, ‘Gene expression analysis’, ‘Cell biology’, ‘Developmental biology’]

Release Date

2017-08-07

Abstract

FACS analysis for E15.5 mouse kidneys protocol by the Potter Group

Procedure

1. Dissect out and isolate E15.5 mouse kidneys in ice-cold PBS.
2. Place four (4) kidney in a 1.5ml eppendorf tube.
3. Add 400ul 0.05% Trypsin-EDTA.
4. Incubate 37oC, five (5) minutes.
5. Triturate 30X.
6. Add 600ul ice-cold 0.1% BSA/PBS or 2% FBS/PBS.
7. Triturate 30X.
8. Centrifuge tubes at 1500 rpm for five (5) minutes. Keep cells cold.
9. Remove supernatant.
10. Resuspend pellet with 600ul ice-cold 0.1% BSA/PBS or 2% FBS/PBS.
11. Recentrifuge tubes at 1500 rpm for five (5) minutes. Keep cells cold.
12. Remove supernatant.
13. Resuspend pellet with 600ul ice-cold 0.1% BSA/PBS or 2% FBS/PBS.
14. Filter cells using 70um mesh FACS collection tube. Keep cells on ice.
15. Immediately FACS using a high-speed digital BD FACS Aria II Cell Sorter.

Consortium

GenitoUrinary Development Molecular Anatomy Project (GUDMAP) Consortium