In dissociated live cells, does staining them with an antibody block the protein's function?

I am sorting cells based on their expression of a few marker genes, and culturing them post-sort for several days.

How can one assess if the protein is still functional?

  • 2
    $\begingroup$ That depends on a number of factors: The actual place where the antibody binds, the protein itself and also the number of available protein molecules. About which protein are you thinking? $\endgroup$
    – Chris
    Commented Jan 12, 2015 at 7:28
  • $\begingroup$ Also, the assay for functionality completely depends on the protein you're testing. Assuming you have an in vitro assay for your protein's function, you could try performing the assay at a given protein concentration with varying concentration of antibody to see if there is a significant effect on activity in vitro. It might not correlate well in vivo, but it is a start. $\endgroup$
    – A. Kennard
    Commented Jan 13, 2015 at 7:34
  • $\begingroup$ Thanks for your input, @Chris. I do not have a specific protein in mind; rather I have a panel of proteins which I want to use to sort cells based on their expression, and I am worried that these antibodies will mess with the cell's behavior, as I will continue to observe them after selecting for them. $\endgroup$ Commented Jan 17, 2015 at 5:17
  • $\begingroup$ Thanks, @A.Kennard. That makes sense, although I don't have an in vitro assay for my proteins' functions. Do you know of any examples where people have verified the activity of proteins for which they added antibodies in vivo? $\endgroup$ Commented Jan 17, 2015 at 5:19


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