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. 2019 Jan 24;20(3):497. doi: 10.3390/ijms20030497

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Identification of serine proteinase inhibitor (SPI) species using biotinylated trypsin and avidin alkaline phosphatase conjugate by affinity blotting in samples from the various isolation procedures used in this study (a) Selected bands were also identified using antibodies to CS (2-B-6 (+)), α1 M, Bikunin, and TSG-6 (b) and the blot intensities summarized in the Table in (c). ITI and pre-α-TI present in ovine plasma (lanes 1, 2, 4, 5) and UTI in urine (lanes 3, 6) were also identified by Western blotting using antibodies to ITI (lanes 1, 2) and bikunin (lane 3) and by affinity blotting using bT (lanes 4–6) (d). The extra band in lane 5 but not in lane 2 in segment (d) is due to α1-proteinase inhibitor which also displays an affinity for HA thus is also included in the pre-α-TI pool taken from Sephadex G200 chromatography in our isolation scheme.