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Updated: Feb 2, 2026

Optimized Analysis of Proteins from Xenopus Oocytes and Embryos by Immunoblotting
Published on: September 19, 2025
Proteolytic Activation of Bmps: Analysis of Cleavage in Xenopus Oocytes and Embryos
Hyung-Seok Kim1, Autumn McKnite1, Jan L Christian2
1Division of Hematology and Hematologic Malignancies, Department of Neurobiology, Anatomy and Internal Medicine, University of Utah School of Medicine, Salt Lake City, UT, USA.
Abstract:
Bone morphogenetic proteins (Bmps) are synthesized as inactive precursors that are cleaved to generate active ligands, along with prodomain fragments that can modulate growth factor activity. Here we provide three protocols that can be used to examine the process of proteolytic activation of Bmps. The first protocol describes how to generate radiolabeled Bmp precursor proteins in Xenopus oocytes and then analyze the time course of precursor cleavage by recombinant enzymes in vitro. The second protocol details how to analyze cleavage of radiolabeled precursor proteins in Xenopus oocytes over time using pulse-chase analysis and autoradiography. This protocol can also be used to analyze folding and cleavage of radiolabeled precursor proteins at steady state. Finally, the third protocol details methods for isolating Bmp cleavage products from the blastocoele of Xenopus embryos and then analyzing them on immunoblots.
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