Despite using published protocols for the use of AMD3100 in mice, is there any way to prove via a quick assay, either immunohistochemical in any mouse tissue or an immunoassay in the circulation, that you are actually achieving the suppression of the CXCR4/CXCL12 pathway? It is fine to show differences in control versus treatment on the phenotype you are investigating, but I guess I am referring to a well-established phenotype that could be used as a positive control to evaluate that your treatments worked.
As an example to what I am asking above, long time ago I used DAPT to inhibit NOTCH pathway in vivo, and I found studies showing that in vivo NOTCH inhibition results in excessive goblet cell differentiation; so ever since, I am always confirming in vivo NOTCH inhibition via a PAS staining in mice duodenum before measuring the phenotype I am interested (my studies are unrelated to GI system - just using the assay as positive control).
Thanks!
This is quite wrong: for ref. see CXCR4 in human (https://pubmed.ncbi.nlm.nih.gov/?term=cxcr4+human) as well as soluble CXCR4 in human sera (e.g. see Malvoisin et al., Anal Biochem. 2011 Jul 15;414(2):202-7. doi: 10.1016/j.ab.2011.03.022).
However, a quick assay seems to be somewhat unrealistic. I would propose to use classical assays for expression (e.g. ELISA, WB, IF etc. - see https://www.rndsystems.com/search?keywords=cxcl12) and function (e.g. see https://www.rndsystems.com/products/human-magnetic-luminex-assay_lxsahm and other R&D products https://www.rndsystems.com/search?keywords=cxcl12).
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