title: Generation of chicken monoclonal antibodies against the a1, a2, and a3 subunit isoforms of vacuolar-type proton ATPase.
abstract: The vacuolar-type proton pump ATPase (V-ATPase) plays several pivotal roles in the acidification of diverse intracellular compartments and the extracellular environment. The a subunit isoforms a1, a2, and a3, constituting the membrane-embedded section, are expressed in various tissues, and they are involved in the regulation of subcellular localization and activity of the holocomplex. Therefore, the characterization of their properties is indispensable for dissection of the physiological roles of the V-ATPase in highly differentiated cells. In this study, we report the production and characterization of chicken monoclonal antibodies (MAbs) against these mouse a1, a2 and a3 subunit isoforms. These MAbs are shown to be suitable for both immunoblotting and immunofluorescence analysis. The MAbs obtained in this study are useful in understanding the pathological basis of V-ATPase dysfunction.
authors.affiliation: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Doshisha Women's College, Kyotanabe, Japan. kwada@dwc.doshisha.ac.jp
authors.forename: Ge-Hong
authors.initials: GH
authors.lastname: Sun-Wada
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tabata
authors.forename: Motoki
authors.initials: M
authors.lastname: Kuhara
authors.forename: Iori
authors.initials: I
authors.lastname: Kitahara
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Takashima
authors.forename: Yoh
authors.initials: Y
authors.lastname: Wada
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D001402:B-Lymphocytes / Q000276:immunology; D015153:Blotting, Western; D002645:Chickens; D003001:Cloning, Molecular; D004926:Escherichia coli; D005260:Female; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D021122:Protein Subunits / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011522:Protons; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D021122:Protein Subunits; D011522:Protons; D011994:Recombinant Proteins; D025262:Vacuolar Proton-Translocating ATPases