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.
Harris
Ceinwen M
CM
Department of Medical Biochemistry and Immunology, University of Wales College of Medicine, Heath Park, Cardiff, Wales CF14 4XN, UK.
Mendes
Filipa
F
Dragomir
Anca
A
Doull
Iolo J M
IJ
Carvalho-Oliveira
I
I
Bebok
Zsuzsanna
Z
Clancy
John P
JP
Eubanks
Valerie
V
Sorscher
Eric J
EJ
Roomans
Godfried M
GM
Amaral
Margarida D
MD
McPherson
Margaret A
MA
Penque
Deborah
D
Dormer
Robert L
RL
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