title: A monoclonal antibody produced against Naked2.
abstract: Naked2 (NKD2) is a member of the Naked family and negatively regulates canonical Wnt signaling. NKD2 may play a role in embryo development and tumor formation by affecting Wnt signaling. In the present study, we describe the establishment of a monoclonal antibody against NKD2 (anti-NKD2 MAb) through the hybridoma method. The purified anti-NKD2 MAb measured a titer of 2.56 × 10(5) against NKD2 by indirect ELISA. Western blot analysis, immunoprecipitation, and confocal microscope showed that the anti-NKD2 MAb can specifically combine NKD2 protein in SW480 and LOVO cells. Competitive inhibition assays of Western blot and indirect ELISA showed that the anti-NKD2 MAb can be blocked with NKD2(1-217) protein. The anti-NKD2 MAb would be helpful for further studies on the structure activity relationship, protein detecting, and cell-signaling pathway of NKD2.
authors.affiliation: Cancer Research Center, Medical College, Xiamen University, Xiamen, Fujian, China.
authors.forename: Chang
authors.initials: C
authors.lastname: Cao
authors.forename: Shengyu
authors.initials: S
authors.lastname: Wang
authors.forename: Sha
authors.initials: S
authors.lastname: Lv
authors.forename: Zhe
authors.initials: Z
authors.lastname: Li
authors.forename: Xianjiang
authors.initials: X
authors.lastname: Wang
authors.forename: Fanwei
authors.initials: F
authors.lastname: Zeng
authors.forename: Haipeng
authors.initials: H
authors.lastname: Zhang
authors.forename: Yujuan
authors.initials: Y
authors.lastname: Dai
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Dou
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Chen
authors.forename: Xiudong
authors.initials: X
authors.lastname: Li
authors.forename: Lingli
authors.initials: L
authors.lastname: Luo
authors.forename: Tianhui
authors.initials: T
authors.lastname: Hu
authors.forename: Jianghua
authors.initials: J
authors.lastname: Yan
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D000917:Antibody Formation; D000918:Antibody Specificity; D015153:Blotting, Western; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007114:Immunization; D047468:Immunoprecipitation; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D011994:Recombinant Proteins; D014407:Tumor Cells, Cultured
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; C449092:NKD2 protein, human; D011994:Recombinant Proteins
references.citation: Mol Cell Proteomics. 2008 Sep;7(9):1651-67
references.pmid: 18504258
references.citation: J Biol Chem. 2010 Apr 30;285(18):13561-8
references.pmid: 20177058
references.citation: Hybridoma (Larchmt). 2011 Aug;30(4):369-73
references.pmid: 21851237
references.citation: Hybridoma (Larchmt). 2012 Jun;31(3):209-13
references.pmid: 22741586
references.citation: Biochem Biophys Res Commun. 2006 Dec 1;350(4):911-5
references.pmid: 17045239
references.citation: Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5571-6
references.pmid: 15064403
references.citation: Mol Biol Cell. 2007 Aug;18(8):3081-93
references.pmid: 17553928
references.citation: Dev Biol. 2007 Sep 15;309(2):151-68
references.pmid: 17689523
references.citation: Cell. 2006 Nov 3;127(3):469-80
references.pmid: 17081971
references.citation: Nature. 1975 Aug 7;256(5517):495-7
references.pmid: 1172191
references.citation: Genes Dev. 1997 Dec 15;11(24):3286-305
references.pmid: 9407023
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 9;105(36):13433-8
references.pmid: 18757723