Abstract
Recombinant expression of secreted Frizzled-related proteins (sFRPs) in mammalian expression systems is a convenient source of these proteins for biological studies. Yields of protein vary; screening of clonal lines for high expression is usually worthwhile. Heparin affinity chromatography is an easy step that provides a major enrichment, particularly for sFRP-1 and sFRP-2. Alternatively, sFRP derivatives tagged with poly-histidine at their carboxyl termini are functional and can be readily isolated by chelating chro-matography. Once purified, the proteins are stable indefinitely if stored frozen and they tolerate multiple rounds of freeze—thawing. Pre-incubation of Wnt samples with sFRP protein for 30 min at 37°C is sufficient to inhibit Wnt activity in various assays. The concentration of sFRP required to block Wnt signaling should be determined empirically, as it will vary with the Wnt preparation and cellular context.
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This research was supported by the Intramural Research Program of the National Institutes of Health, National Cancer Institute.
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Wolf, V., Endo, Y., Rubin, J.S. (2008). Purification and Wnt-Inhibitory Activities of Secreted Frizzled-Related Proteins. In: Vincan, E. (eds) Wnt Signaling. Methods in Molecular Biology™, vol 468. Humana Press. https://doi.org/10.1007/978-1-59745-249-6_3
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DOI: https://doi.org/10.1007/978-1-59745-249-6_3
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