I am measuring phosphorylated STAT5 in different lymphocyte populations using flow cytometry in response to stimulus. I need to accurately gate different memory populations using CD27 (not CCR7, as I use cryopreserved material).
Can you please recommend an antibody-fluorochrome combination proven to work with BD Phosflow buffer III (an 80% methanol-based buffer)?
The BD Cytobank Buffer compatibility tool ( https://www.cytobank.org/facselect/ )suggests clone CD27 L128, but the fluorochromes are not suitable for my needs as these channels are already occupied (APC, PE and FITC). Has anyone tested a more extensive range of fluorochromes for this marker and technique please?
You basically need CD27 (L128) which efficiently work with phosflow buffer III. In the cytobank app you can only find fluorochromes for channels which you already have occupied. So you should now search for another fluorochrome which works well with buffer-III. I searched and found that CD27 (L128) is available in PerCP-Cy5.5 which you have an option. Then searched for it's (PerCP-Cy5.5) compatibility with phosflow BUFFER-III, AND found a pSTAT-5 in same fluorochrome. It means if you choose CD27 (L128) in PerCp-Cy5.5 (cat no. 656643) it would work with phosflow BUFFER-III and fits in your panel.
Attaching evidence for the same.
You basically need CD27 (L128) which efficiently work with phosflow buffer III. In the cytobank app you can only find fluorochromes for channels which you already have occupied. So you should now search for another fluorochrome which works well with buffer-III. I searched and found that CD27 (L128) is available in PerCP-Cy5.5 which you have an option. Then searched for it's (PerCP-Cy5.5) compatibility with phosflow BUFFER-III, AND found a pSTAT-5 in same fluorochrome. It means if you choose CD27 (L128) in PerCp-Cy5.5 (cat no. 656643) it would work with phosflow BUFFER-III and fits in your panel.
Attaching evidence for the same.
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