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. 2013 Mar 29;8(3):e60638. doi: 10.1371/journal.pone.0060638

Figure 3. Proteins extracted from PFPE tissues are suitable for ELISAs with antibodies directed against denatured proteins.

Figure 3

Three human malignant (breast cancer metastasis, ovarian and prostate cancer) and three non-malignant (stomach, duodenum, uterus) tissue specimens were each divided into two samples and either cryopreserved (cryo) or fixed and stabilized in the PAXgene Tissue reagents and paraffin-embedded (PFPE). Proteins were extracted with respective protocols (See ELISA, Experimental Section) using denaturing or non-denaturing extraction buffer. Results are always depicted in duplicates for each lysate. (A) An ELISA for non-denatured Akt (n = 2 for stomach and duodenum samples. (C) An ELISA for denatured Erk-1/2 (n = 2). (E) An ELISA for denatured phospho-Erk-1/2 (n = 3). Western blot analysis of (B) 12.5 µg or (D, F) 25 µg protein of each lysate separated by SDS-PAGE and performed using indicated antibodies.