I'm doing conceptual questions in preparation for a Western Blott and needed clarification on a specific question:

Is it possible/ will your Western blot work if you use a primary antibody (say generated in mice) and use a secondary antibody (generated in goats)? Would this combination still work with the western blot? At first glance, I'd say no but I'm still looking into it. Any clarification would be greatly appreciated!

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    $\begingroup$ It's not clear what you're asking, but a typical Western would indeed involve a primary mouse antibody and a secondary goat anti-mouse antibody; the primary would bind to your targets, the secondary would have the detector molecule attached and would bind to the primary. $\endgroup$ – iayork Mar 22 '18 at 19:20

As others pointed out, the question is not clear.. but yes, in general you use a primary antibody recognizing your peptide of interest, e.g. an antibody made in mouse, and then a secondary antibody recognizing your primary antibody, in this case an anti-mouse. The secondary antibody will obviously be made in a different animal than mouse (e.g. in goat), because mice cannot develop anti-mouse antibodies! I hope I understood your question.

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