Assessment of CFTR localisation in native airway epithelial cells obtained by nasal brushing.
Reliable methods for determining the localisation of mutant CFTR protein in native cells from CF individuals are necessary to allow the degree of mislocalisation of any genotype to be defined and to assess the effect of therapeutic agents on CFTR trafficking. Here, we present procedures for obtaining ciliated epithelial cells from CF patients by nasal brushing and a description of protocols for immunolocalisation of CFTR. The protocols are a consensus, following comparison of some aspects of methods currently used in the authors' laboratories.
Department of Medical Biochemistry and Immunology, University of Wales College of Medicine, Heath Park, Cardiff, Wales CF14 4XN, UK.
Ceinwen M
CM
Harris
Filipa
F
Mendes
Anca
A
Dragomir
Iolo J M
IJ
Doull
I
I
Carvalho-Oliveira
Zsuzsanna
Z
Bebok
John P
JP
Clancy
Valerie
V
Eubanks
Eric J
EJ
Sorscher
Godfried M
GM
Roomans
Margarida D
MD
Amaral
Margaret A
MA
McPherson
Deborah
D
Penque
Robert L
RL
Dormer
D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000302:isolation & purification*; D004847:Epithelial Cells / Q000737:chemistry; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D020545:Respiratory Mucosa / Q000737:chemistry / Q000473:pathology*; D013048:Specimen Handling / Q000379:methods
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator