Well, if the protein you introduce is identical to the endogenously expressed one, then you will not be able to distinguish between the two. If you simply want to check whether your expression constuct is working at all, you can do western blot analysis and compare your transfected cells to a non-transfected control. Since your plasmid is designed for overexpression, the amount of target protein you can detect in your cell lysates should increase dramatically after transfection with your plasmid.

Nonetheless, I would consider adding a small tag (Flag, Myc, etc.) to your plasmid. It is a rather easy procedure and will allow you to use tag-specific antibodies to distiguish between endogenous and recombinant proteins.

Good Luck!

Go for western blot analysis using primary antibody against your protein or against any epitopes you have fused with your protein of interest.

There is no tag fused with my protein of insert. I was given with the plasmid pcdna3. 1 having no tag. I have inserted my gene of interest into that plssmid and made a protein. As there is no tag so now how can i detect the exogenous protein. @Yeshveer Singh

Well, if the protein you introduce is identical to the endogenously expressed one, then you will not be able to distinguish between the two. If you simply want to check whether your expression constuct is working at all, you can do western blot analysis and compare your transfected cells to a non-transfected control. Since your plasmid is designed for overexpression, the amount of target protein you can detect in your cell lysates should increase dramatically after transfection with your plasmid.

Nonetheless, I would consider adding a small tag (Flag, Myc, etc.) to your plasmid. It is a rather easy procedure and will allow you to use tag-specific antibodies to distiguish between endogenous and recombinant proteins.

Good Luck!

@Ramal Jahid

In such case, do a simple comparative SDS-PAGE analysis with transfected and non-transfected control as mentioned by @Jan Philipp Robert. You will see increased intensity of corresponding band, if your protein is expressing.

Dear Ramal

i'm agree with Jan Philipp.

If you are expressing the full length protein, and the protein is human and expressed in human cell lines, probably you will not able to distinguish the 2 forms if you are not add a tag (also an His-tag could be useful) to the exogenous version.

I'm not expert about P53 and i do not know how much is the homology between the human and hamster protein homologues but in case that there are some differences and you can find antibodies specific for those different forms you can try to express you protein with the CHO.

good luck

Manuele