Hello, I have observed a strange result after running IHC and western blot. I have determined the expression of my protein in a tumor cell line on the one hand with immunohistochemistry (ihc) and on the other hand with a sds-page/western blot.
The result was that I got a nice signal on the ihc but I failed with the WB. It´s very strange to me because of using the same primary antibody for both techniques. What´s the reason for this phenomenon? Which result is reliable? How can I verify these results? I am really helpless. Regards Tiffy
PS The primary ab works well on another cell line in the wb.

Hi Tiffy,

Can you give us some more information on your experiments?

How many times have you repeated the IHC and Western blot? Did all your controls work correctly? What were the controls? My guess is the protein was somehow denatured in the western blot and no longer sees the antibody. The other possibility is the antibody has somehow gone bad on you.

Also what is the evidence that the western blot with the "other" cell line gives you the correct size band?

Rb