Journal of Clinical Endocrinology & Metabolism, Vol 72, 1391-1393, Copyright © 1991 by Endocrine Society

ARTICLES

A simple assay for proteolysis of IGFBP-3

G Lamson, LC Giudice and RG Rosenfeld
Department of Pediatrics, Stanford University Medical Center, California 94305.

An assay for assessing the proteolysis of IGFBP-3 by a biological fluid or protein has been developed using 125IhIGFBP-3 recombinantly derived from Escherichia coli as a substrate indicator. The labelled substrate is incubated at 37 degrees in the presence of the agent being tested and proteolysis is visualized by SDS-PAGE and autoradiography. The assay showed that the pregnancy associated protease is Ca++ dependent. The substrate is also proteolyzed by rat and mouse pregnancy sera, making the assay applicable to studies in those animal systems.

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