If it is necessary for EGFR to have its native structure in order to be detected by Western blot, the antibody you are using must recognize a structural epitope. Is there another antibody you could use that recognizes a continuous epitope? Then you could use a traditional method of Western blotting, starting with SDS-PAGE.

If I remember it correctely we blotted EGFR and Phospho-EGFR like any other protein in a practical training course during academic studies. Did you ever try?

You may need EGFR in native form to bind EGF but not with many anti-EGFR antibodies. So if your objective is to detect EGFR expression, then just try to use another anti-EGFR antibody that detect EGFR in denature form. This will solve your problems with native gel electrophoresis. Alternatively use ELISA to measure EGFR expression. If EGFR is heat sensitive, why are heating it at 75C? If you are using membrane fraction or whole cells as the sample for EGFR detection, first solubilise the sample in appropriate detergent (NP-40 or Triton X-100), mix soluble protein with 1/10th volume of 50% sucrose and 0.1% bromophenol blue and then load in native PAGE (make sure there is no SDS in any electrophoresis buffers). Also run gel at 4C to avoid denaturing of EGFR. Although I never used it myself, I have seen many people recommending blue native PAGE for native electroblotting and immunodetection (http://www.nature.com/nprot/journal/v1/n1/full/nprot.2006.62.html).

Thanks,

Do you know the problem, it lasts about  four to five weeks in order to receive any antibody, for that I keep trying and trying, for 75 C  I read about that in an article they said it will increase yield up to 79 C, I know that EGFR is a heat sensitive