We have begun doing experiments with human tissue and have the following markers:
CD45
DAPI
------
CD3
CD4
CD8
CD11b
CD11c
We routinely use CD45 for lymphocytes and DAPI for live cell selection. CD3/CD4+ or CD8+ obviously we can use to detect these types of T cells but I find alot of ambiguity with CD11b and CD11c. What specific cell types can I detect in human tissue with these Ab's (including gating on CD3- populations)?
Thanks in advance!
CD4+ T cells, CD8+ T cells, CD11c+ DCs can be distingushed by this penel.
When you say staining in tissue, do you mean flow staining or histology? I'm going to assume flow.
CD45 stains all leukocytes, so why are you using these for only lymphocytes? I"m goin to assume that you are gating on the lymphocyte population by the SSC?
CD11c as a DC marker works better in mouse, but it is usually useful in humans... but what tissue are you looking at? You might be able to find a better marker.
As for CD11b, this is express on a lot of leukoyctes. (It's expression isn't high or uniform on lymphocytes, which is why it's usually used as a monocyte/macrophage marker). But to use CD11b, it usually involves either using a secondary marker (such as CD66b to rule out neutrophils) or you use the FSC/SSC plot to gate out the macrophage/monocytes and CD11b is usually used more for confirmation of that gating strategy.
One more thing..to find CD11c and CD11b, you can't just gate on CD3- cells especially if you are gating on the lymphocyte cluster by SSC. Most cells that express these markers do not fall into the lymphocyte cluster.
Dear Chris Cook
CD3 is an essential T cell co-receptor and defines T cell lineage.
Best wishes,
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology