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. 2015 Aug 14;10(8):e0135374. doi: 10.1371/journal.pone.0135374

Fig 3. PN extracted from fibrotic human lung has the pI expected of un-modified protein and does not react with anti-Gla.

Fig 3

Approximately 200 μg of protein extracted with SDS from fibrotic lung samples were subjected in parallel to 2-dimensional isoelectric focusing/electrophoresis, transferred to PVDF membranes, and Western blotted. The membranes were stained with Coomassie blue (A and B) and probed with (C) mouse anti-PN or (D) mouse anti-Gla mAbs. Enlarged views of the PN-positive region are shown in the insets for both blots. Isoelectric points of PN-positive spots ranged between pH 7.0 and > pH 8.0. Gla-positive spots were more acidic and did not overlap with PN positive spots. An internal isoelectric focusing standard, tropomyosin, migrated as a doublet with the lower polypeptide spot (arrows) of MW 33 kDa and pI 5.2 on the parallel 10% gels.