I am putting together a panel for flow cytometry, for one particular antibody the only available isotype control has a different light chain to the test antibody. My test antibody is BV711-CD25 Rat IgG1 lambda... the only isotype control available was BV711 Rat IgG1 kappa.
Would this still be OK too use as the isotype control, or do kappa/lambda chains have to be the same?
It's very rare to ever need to use an isotype control for flow anymore.
But if you really want to...yes, it should be fine. Because you are technically trying to detect nonspecific binding to the Fc receptors, so the light chains shouldn't matter...
Harry,
Most of the antibody binding receptors on the surface of the cell recognize the Fc regions of antibodies that are made of heavy, not light chains (like Fc gamma receptors, for example). So to those receptors it does not matter what the nature of the light chain is. You control is good enough. You can, of course, use Fc blocker to decrease the background signal - especially if you don't fix and permeabilize your cells. Good luck!
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