Please use this identifier to cite or link to this item: https://repositorio.uca.edu.ar/handle/123456789/14595
Título : Measurement of cystic fibrosis transmembrane conductance regulator activity using fluorescence spectrophotometry
Autor : Valdivieso, Ángel Gabriel 
Marín, María C. 
Clauzure, Mariángeles 
Santa Coloma, Tomás Antonio 
Palabras clave : FIBROSIS QUÍSTICACANAL DE CLORUROESPECTROFOTOMETRÍAREGULADOR DE CONDUCTANCIA TRANSMEMBRANA DE LA FIBROSIS QUÍSTICAFLUORESCENCIA
Fecha de publicación : 2011
Editorial : Elsevier
Cita : Valdivieso, Á. G. Measurement of cystic fibrosis transmembrane conductance regulator activity using fluorescence spectrophotometry [en línea]. Analytical Biochemistry Bioquímica Analítica. 2011, 418 (2). doi: 10.1016/j.ab.2011.07.029. Disponible en: https://repositorio.uca.edu.ar/handle/123456789/14595
Resumen : Abstract: Cystic fibrosis (CF) is a frequent autosomal recessive disease caused by mutations that impair the CF transmembrane conductance regulator (CFTR) protein function. CFTR is a chloride channel activated by cyclic AMP (cAMP) via protein kinase A (PKA) and ATP hydrolysis. We describe here a method to measure CFTR activity in a monolayer of cultured cells using a fluorescence spectrophotometer and the chloridesensitive probe 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ). Modifying a slice holder, the spectrophotometer quartz cuvette was converted in a perfusion chamber, allowing measurement of CFTR activity in real time, in a monolayer of T84 colon carcinoma cells. The SPQ Stern–Volmer constant (KCl ) for chloride in water solution was 115.0 ± 2.8 M1 , whereas the intracellular KCl was 17.8 ± 0.8 M1 , for T84 cells. A functional analysis was performed by measuring CFTR activity in T84 cells. The CFTR transport inhibitors CFTR(inh)-172 (5 lM) and glibenclamide (100 lM) showed a significant reduction (P < 0.05) in CFTR activity. This simple method allows measuring CFTR activity in a very simple, reproducible, and sensitive way.
URI : https://repositorio.uca.edu.ar/handle/123456789/14595
ISSN : 0003-2697
1096-0309 (online)
Disciplina: MEDICINA
DOI: 10.1016/j.ab.2011.07.029
Derechos: Acceso restringido
Appears in Collections:Artículos

Files in This Item:
File Description SizeFormat Existent users please Login
measurement-cystic-fibrosis-transmembrane.pdf615,9 kBAdobe PDF???org.dspace.app.webui.jsptag.ItemTag.embargo???    Request a copy
Show full item record

Page view(s)

61
checked on Apr 20, 2024

Download(s)

13
checked on Apr 20, 2024

Google ScholarTM

Check


Altmetric


This item is licensed under a Creative Commons License Creative Commons