Total bone marrow preparation
Lineage depletion
LKS or long term HSC sorting
Add antibodies:
Sca PE-Cy5.5 | 1: 100 | 10ml |
Kit APC | 1:100 | 10ml |
SA PE-Cy7 | 1:500 | 2ml |
To sort long term HSC, aslo add:
CD48 FITC | 1:1000 | 1ml |
Flk2 PE | 1:200 | 5ml |
Also, prepare single staining controls using the total bone marrow kept aside after the dissection and the total bone marrow lineage stained. These stainings can be done directly into FACS tubes.
Make sure to be in the dark!