I wish to detect a protein of interest by combing direct and indirect staining for flow cytometry. The idea would be to stain mouse splenocytes and lymph node cells with fluorochrome-conjugated antibodies against surface lineage markers, and then detect my protein of interest via indirect staining.
Would this approach be feasible?
I would reverse the order!!! First indirect and then direct (fluorochrome-conjugated antibodies); Otherwise you should use antibodies of different species (beeing the secondary antibody not cross-reactive).
Yes it is possible Just do it first the direct and then indirect
Best
I would reverse the order!!! First indirect and then direct (fluorochrome-conjugated antibodies); Otherwise you should use antibodies of different species (beeing the secondary antibody not cross-reactive).
Thanks for the suggestions. The protein of interest is a transmembrane protein. The antibody is directed against an extracellular epitope of the protein. Thankfully, no intracellular staining is required...
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