If the protein you are expressing is human, the antibody should be anti human antibody to detect it.

Yes ma‘am. What if it‘s expressing in bacteria? Does it make a difference ? I actually thought of tagging the protein with a Flag tag and use an anti Flag antibody

Dear Lakshami

the main differences regards the post translational modification . eg glycosilation thay may happen if your protein is expressed in mammalian cells but not in e.coli.

regarding the antibody are you speak about a policlonal or a monoclonal?

policlonal contain a a lot of antibodies recognizing different epitopes that i'm quite sure that also if produced kg immumizing protein produced in mammalian there is very high probability thay can recognize also you form from e.coli.

for a monoclonal this is not obvious hoeever the fact that is able to recognize the human form do not mean that it was generated by immunitation of the human protein expressed or extracted from mammalian cells but it could be generated also from the protein produced in e.coli or by peptide immunization and just validate for be used in wb.

Of course if the anotibody was produced by using as imminogen the protein expressed from e.coli the probability that it is recognize also your are higher. You have to try.

good luck

Manuele