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J Immunol Methods. 2010 Jan 31;352(1-2):153-60. doi: 10.1016/j.jim.2009.11.016. Epub 2009 Nov 26.

Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation.

Author information

1
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.

Abstract

LRP6 is a cell surface molecule that plays a critical role in the Wnt signaling pathway, and is implicated in numerous human diseases. Studies of cellular signaling mediated by LRP6 have relied on overexpression experiments, due to the lack of good monoclonal antibodies (mAbs) reactive with native LRP6 ectodomain. By using native recombinant LRP6 ectodomain fragment produced in mammalian expression system, we succeeded in developing a panel of anti-human LRP6 mAbs. Selected mAbs were capable of staining endogenous LRP6 on cell surface by using flow cytometry and immunofluorescence microscopy, and enriching detergent-solubilized LRP6 from cell lysate by immunoprecipitation.

PMID:
19945460
DOI:
10.1016/j.jim.2009.11.016
[Indexed for MEDLINE]

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