CFTR Protein Immunoprecipitation and Detection

Whole-cell lysates were prepared as described previously (74 (link)). CFTR was immunoprecipitated as described previously (35 (link), 73 (link)) and isolated using Protein A/G PLUS agarose (Santa Cruz Biotechnology). Samples were separated on 4–20% gradient SDS-PAGE gels (Bio-Rad) and then transferred to nitrocellulose. Blots were probed with mouse monoclonal anti-CFTR antibodies and then with IR Dye 680-goat anti-mouse IgG (Molecular Probes). Anti-actin (Cell Signaling) or anti-tubulin (LI-COR) was used as a loading control. Protein bands were visualized using an Odyssey Infrared Imaging System (LI-COR).

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Cholon D.M., Quinney N.L., Fulcher M.L., Esther CR J.r., Das J., Dokholyan N.V., Randell S.H., Boucher R.C, & Gentzsch M. (2014). Potentiator Ivacaftor Abrogates Pharmacological Correction of ΔF508 CFTR in Cystic Fibrosis. Science translational medicine, 6(246), 246ra96.

Publication 2014

Protein A/G PLUS agarose 4–20% gradient SDS-PAGE gels Anti-actin anti-tubulin Odyssey Infrared Imaging System

Actin Agarose Anti igg Antibodies Cell Cftr Cftr mouse Gels Goat Molecular probes Mouse Nitrocellulose Protein Protein a Sds page Tubulin

Corresponding Organization : Lung Institute

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Variable analysis

independent variables

Whole-cell lysates were prepared as described previously (74 (link)).
CFTR was immunoprecipitated as described previously (35 (link), 73 (link)) and isolated using Protein A/G PLUS agarose (Santa Cruz Biotechnology).

dependent variables

Protein bands were visualized using an Odyssey Infrared Imaging System (LI-COR).

control variables

Anti-actin (Cell Signaling) or anti-tubulin (LI-COR) was used as a loading control.

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