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Titolo:
Detection and quantitation of a sodium-dependent sulfate cotransporter (NaSi-1) by sandwich-type enzyme-linked immunosorbent assay
Autore:
Sagawa, K; DuBois, DC; Han, B; Almon, RR; Biber, J; Murer, H; Morris, ME;
Indirizzi:
SUNY Buffalo, Dept Pharmaceut, Amherst, NY 14260 USA SUNY Buffalo AmherstNY USA 14260 Dept Pharmaceut, Amherst, NY 14260 USA SUNY Buffalo, Dept Biol Sci, Amherst, NY 14260 USA SUNY Buffalo Amherst NY USA 14260 o, Dept Biol Sci, Amherst, NY 14260 USA Univ Zurich, Inst Physiol, CH-8057 Zurich, Switzerland Univ Zurich Zurich Switzerland CH-8057 siol, CH-8057 Zurich, Switzerland
Titolo Testata:
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY
fascicolo: 1, volume: 437, anno: 1998,
pagine: 123 - 129
SICI:
0031-6768(199812)437:1<123:DAQOAS>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
BORDER MEMBRANE-VESICLES; INORGANIC SULFATE; BASOLATERAL MEMBRANE; DIETARY SULFATE; RAT-KIDNEY; TRANSPORT; EXPRESSION; ADAPTATION; INHIBITION; CELLS;
Keywords:
sodium-dependent sulfate cotransport; rat kidney cortex; antibodies; immunoassay; ELISA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
28
Recensione:
Indirizzi per estratti:
Indirizzo: Morris, ME SUNY Buffalo, Dept Pharmaceut, Amherst, NY 14260 USA SUNY Buffalo Amherst NY USA 14260 aceut, Amherst, NY 14260 USA
Citazione:
K. Sagawa et al., "Detection and quantitation of a sodium-dependent sulfate cotransporter (NaSi-1) by sandwich-type enzyme-linked immunosorbent assay", PFLUG ARCH, 437(1), 1998, pp. 123-129

Abstract

The sodium-dependent sulfate transporter (NaSi-1) DNA has been recently identified from rat kidney cortex. The objective of this study was to developa quantitative assay for the NaSi-1 transporter protein. The NaSi-1 antigen was prepared by fusion protein techniques following analysis of the primary sequence for antigenicity. Polyclonal and monoclonal antibodies against the NaSi-1 antigen were raised in rabbits and mice, respectively. The specificity of the raised antibodies was examined by Western analysis using brush-border membrane (BBM) and basolateral membrane (BLM) purified from rat kidney cortex. Both NaSi-1 polyclonal and monoclonal antibodies detected a 69-kDa protein in the BBM. Using the purified monoclonal antibody as the capture antibody and the polyclonal antibody as the detecting antibody, a simple and sensitive sandwich-type enzyme linked immunosorbent assay was developed to quantitate NaSi-1 transporter protein levels in tissue. The specificity of the assay was examined using BBM, BLM and NaSi-1-transfected Madin-Darby canine kidney cells. The assay was capable of detecting NaSi-1 at levels as low as 6.58 fmol. The concentration of NaSi-1 transporter protein in crude membrane isolated from rat kidney cortex was 0.094+/-0.014 fmol/mu g protein (mean+/-SD of three preparations).

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Documento generato il 02/07/20 alle ore 18:55:30