Protocol

Cystic Fibrosis

Volume 742 of the series Methods in Molecular Biology pp 15-33

Date:

Imaging CFTR Protein Localization in Cultured Cells and Tissues

  • Silvia M. KredaAffiliated withCystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill
  • , Martina GentzschAffiliated withCystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina at Chapel HillDepartment of Cell and Developmental Biology, University of North Carolina at Chapel Hill Email author 

* Final gross prices may vary according to local VAT.

Get Access

Abstract

CFTR functions as a chloride channel at the apical membrane of airway, gastrointestinal, and other epithelial cells. Immunofluorescence microscopy is commonly used to assess the subcellular localization and relative abundance of CFTR. Visualization of heterologously overexpressed CFTR is typically unproblematic and straightforward, whereas detection of small quantities of endogenous CFTR in tissues can be challenging and requires highly specific antibodies and optimized staining protocols. CFTR tagged by green fluorescent protein can be employed to study trafficking in live cells. Tagging of CFTR with an extracellular epitope permits detection exclusively at the cell surface and subsequent chasing allows visualization of endocytic trafficking.

Key words

CFTR immunostaining CFTR expression CFTR localization airway epithelium F508del-CFTR