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The sodium-dependent inorganic phosphate transporter SLC34A1 (NaPi-IIa) is not localized in the mouse brain: a case of tissue-specific antigenic cross-reactivity.
Department of Anatomy and Centre for Molecular Biology and Neuroscience, University of Oslo, Oslo, Norway; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.ORCID iD: 0000-0003-3584-7829
Department of Anatomy and Centre for Molecular Biology and Neuroscience, University of Oslo, Oslo, Norway.
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, México D.F., Mexico.
Institute of Physiology, University Zürich, Zürich, Switzerland.
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2011 (English)In: Journal of Histochemistry and Cytochemistry, ISSN 0022-1554, E-ISSN 1551-5044, Vol. 59, no 9, 807-812 p.Article in journal (Refereed) Published
Abstract [en]

The sodium-dependent inorganic phosphate transporter NaPi-IIa is expressed in the kidney. Here, the authors used a polyclonal antiserum raised against NaPi-IIa- and NaPi-IIa-deficient mice to characterize its expression in nervous tissue. Western blots showed that a NaPi-IIa immunoreactive band (~90 kDa) was only present in wild-type kidney membranes and not in kidney knockout or wild-type brain membranes. In the water-soluble fraction of wild-type and knockout brains, another band (~50 kDa) was observed; this band was not detected in the kidney. Light and electron microscopic immunohistochemistry using the NaPi-IIa antibodies showed immunolabeling of kidney tubules in wild-type but not knockout mice. In the brain, labeling of presynaptic nerve terminals was present also in NaPi-IIa-deficient mice. This labeling pattern was also produced by the NaPi-IIa preimmune serum. The authors conclude that the polyclonal antiserum is specific toward NaPi-IIa in the kidney, but in the brain, immunolabeling is caused by a cross-reaction of the antiserum with an unknown cytosolic protein that is not present in the kidney. This tissue-specific cross-reactivity highlights a potential pitfall when validating antibody specificity using knockout mouse-derived tissue other than the specific tissue of interest and underlines the utility of specificity testing using preimmune sera.

Place, publisher, year, edition, pages
Sage Publications, 2011. Vol. 59, no 9, 807-812 p.
Keyword [en]
electron microscopy, immunofluorescence, polyreactive, polyclonal, specificity
National Category
Neurosciences
Identifiers
URN: urn:nbn:se:liu:diva-133050DOI: 10.1369/0022155411411713ISI: 000294070400001PubMedID: 21606201Scopus ID: 2-s2.0-80054029153OAI: oai:DiVA.org:liu-133050DiVA: diva2:1054578
Available from: 2016-12-08 Created: 2016-12-08 Last updated: 2016-12-13Bibliographically approved

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