Dear All, Sorry to be disturbing you all with my inquiry. I am tagging a membrane protein (MHC I) just after the Signal peptide with either HA-tag or Myc-tag and expressing it in mammalian cells. I am able to detect that protein by immunoprecipitation followed by Western blotting using anti-Myc antibody or anti-HA antibody. However, I could not detect it by flowcytometry, which could mean the tag was cleaved off or the protein is incapable to be delivered to the cell surface among other possibilities . Does anyone has any experience with tagging membrane proteins after the Signal peptide? Is there any restrictions against that? Or a better tag to use? Thank you.Best regards,Samer
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