Hi
I'm trying to study T-cell differentiation using a protocol from a paper. On the panel the authors have used anti-CD27–PE-Cy7 (clone 1A4CD27; Beckman Coulter) and anti-CCR7–FITC (clone 150503; BD Pharmingen). I want to purchase all my antibodies from biolegend. The available clones from biolegend for anti-CD27 are O323, M-T271 and QA17A18 and for anti-CCR7 the clone is G043H7.
Do you have any experience working with any of these clones?
thanks
mostafa
Hi, I don't know if you should feel restricted to these markers let alone these clones, there are many ways to skin a cat. I expect what type of differentiation you want is a factor but for CD4 I would consider; CD4, CD127, CD25, CD183 and CD196. This will give you Th1, Th2, Th17 and putative Treg. These are all available from Biolegend I expect.
anti-CCR7 the clone is G043H7 is the best clone in my opinion, but don't use a dim fluorochrome as FITC. BV412, PE or APC will give you a much better separation of your cell populations. Please use other markers such as CD45RA, CD45RO and CD62L to better differentiate naive/memory populations. Choosing a specific clone is less critical for CD27 staining.
The anti-CD27 clone O323 works good for human CD4 and CD8 T cells. I have also only used the CCR7 clone 150503 from BD (as the authors in your paper), which worked equally well for us. Including more differentiation markers will definately increase the accuracy of your dataset.
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