Thermo Fisher Scientific (Invitrogen)
Podoplanin Monoclonal Antibody (eBio8.1.1 (8.1.1)), PerCP-eFluor™ 710, eBioscience
46-5381-82
N/A
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We are interested in studying the mechanisms responsible for ensuring T cell self tolerance that rely on thyme and extra-thymic Aire expressing cells.
Flow Cytometry
Mouse thymus
Live dead: Zombie NIR LD 1:4000 in 100ul FACS for 20 min at RT
2.4G2
Surface stains: Pdpn 1:50 in 100uL FACS for 30 min at 4C
PFA (15 min RT) + FoxP3 Fix/Perm kit (30 min 4C), then intracellular stain in 100uL perm buffer for 1hr at RT
Cytek Aurora spectral flow cytometry
This antibody gives decent staining to separate immature mTECs from mature/post-Aire mTECs after titration. However, manufacturer supplied protocol gives only weak detection of Pdpn+ cells by spectral flow, requiring a higher than usual concentration.
Can give good separation of positive and negative populations.
Requires optimization to work on mTECs.
Useful antibody for broadly separating mature and immature mTEC populations.