Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation.
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.
Yasui
Norihisa
N
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Mihara
Emiko
E
Nampo
Maiko
M
Tamura-Kawakami
Keiko
K
Unno
Hideaki
H
Matsumoto
Kyoichi
K
Takagi
Junichi
J
D000818:Animals; D000911:Antibodies, Monoclonal; D002469:Cell Separation; D000939:Epitopes; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D026502:LDL-Receptor Related Proteins / Q000819:agonists / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D060488:Low Density Lipoprotein Receptor-Related Protein-6; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D011487:Protein Conformation; D017434:Protein Structure, Tertiary / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D051822:T Cell Transcription Factor 1 / Q000378:metabolism; D014162:Transfection; D051153:Wnt Proteins / Q000494:pharmacology; D060508:Wnt3 Protein
D000911:Antibodies, Monoclonal; D000939:Epitopes; D026502:LDL-Receptor Related Proteins; C555475:LRP6 protein, human; D060488:Low Density Lipoprotein Receptor-Related Protein-6; C555476:Lrp6 protein, mouse; D011994:Recombinant Proteins; D051822:T Cell Transcription Factor 1; D051153:Wnt Proteins; D060508:Wnt3 Protein