R&D Systems
Mouse CCL21/6Ckine Alexa Fluor® 405-conjugated Antibody
IC457V-100UG
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: 1:100 in 100uL FACS for 30 min at 4C
PFA (15 min RT) + FoxP3 Fix/Perm kit (30 min 4C), then intracellular stain for Ccl21 1:100 in 100uL perm buffer for 1hr at RT
Cytek Aurora spectral flow cytometry
This antibody gives strong separation of Ccl21+ MHCII low immature mTECs (mTEC lo) from the MHCII high mature mTECs and post-Aire mTECs in the thymus. Great for sorting/enriching for desired mTEC sub populations in flow cytometry experiments using spectral flow. Does not require much titration to get optimal staining, manufacturer recommended protocol works well.
Great staining of immature TECs.
No downsides.
Great antibody to use for mTEC studies.