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. 2016 Nov 28;7:542. doi: 10.3389/fimmu.2016.00542

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Copyright © 2016 Schäfer, Grosche, Reinders, Hauck, Pouw, Kuijpers, Wouters, Ehrenstein, Enzmann, Zipfel, Skerka and Pauly.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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MAb RETC-2 detected native FHR-3 in complex with other complement proteins from human serum. (A) MAb RETC-2 detected four protein bands for FHR-3-glycoforms between 35 and 65 kDa (arrows) from reduced normal human serum (NHS) in Western blot. (B) More than 30 different proteins were immunoprecipitated from non-reduced NHS using RETC-2 and isotype control (not shown). Labeled fractions after Coomassie staining were identified by mass spectrometry analyses and contained FHR-3. Unlabeled protein bands contained other serum proteins (e.g., serum albumin, apolipoproteins, Igs). (C) RETC-2 specifically precipitated FHR-3 (green star) in comparison to an isotype control mAb. Additional peptide fragments, characterized with non-gel LC-MS/MS, were complement proteins.