Characterization of CFTR Expression: A Methodological Comparison of SDS-PAGE Systems
Zoom Link
https://muw.zoom.us/j/98287107796
Department
Sciences & Mathematics
Format of Presentation
Oral Presentation
Research Category
STEM
Description
CFTR functions as a chloride ion channel on the plasma membrane of many epithelial cells. Mutations alter the function of CFTR, leading to diseases such as cystic fibrosis, cholera, and diarrhea. Since CFTR expression levels are negligible in native cells, gene transfection is necessary to overexpress this protein, which can then be easily detected by western blotting, a widely used biochemical method for protein detection. The goal of this study is to determine the optimal electrophoretic conditions for detecting CFTR expression levels. Protein samples from a lung epithelial cell line stably transfected with either wild-type or mutated (ΔF508) CFTR gene were prepared and electrophoresed under native or denaturing conditions using continuous or discontinuous SDS-PAGE buffer systems. Discontinuous SDS-PAGE produced sharper, more defined bands compared to the continuous system. CFTR expression is more enhanced under denaturing conditions than native conditions. However, boiling the samples under denaturing conditions distorts the signals. These findings may help researchers identify suitable conditions for detecting expression levels of CFTR and other proteins.
Recommended Citation
Tasnim Raya, Zarin; Wilkins, Ethan R.; and Heda, Ghanshyam, "Characterization of CFTR Expression: A Methodological Comparison of SDS-PAGE Systems" (2026). Undergraduate Research Conference. 2.
https://athenacommons.muw.edu/urc/2026/stem-and-social-sciences-oral-presentations/2
Characterization of CFTR Expression: A Methodological Comparison of SDS-PAGE Systems
CFTR functions as a chloride ion channel on the plasma membrane of many epithelial cells. Mutations alter the function of CFTR, leading to diseases such as cystic fibrosis, cholera, and diarrhea. Since CFTR expression levels are negligible in native cells, gene transfection is necessary to overexpress this protein, which can then be easily detected by western blotting, a widely used biochemical method for protein detection. The goal of this study is to determine the optimal electrophoretic conditions for detecting CFTR expression levels. Protein samples from a lung epithelial cell line stably transfected with either wild-type or mutated (ΔF508) CFTR gene were prepared and electrophoresed under native or denaturing conditions using continuous or discontinuous SDS-PAGE buffer systems. Discontinuous SDS-PAGE produced sharper, more defined bands compared to the continuous system. CFTR expression is more enhanced under denaturing conditions than native conditions. However, boiling the samples under denaturing conditions distorts the signals. These findings may help researchers identify suitable conditions for detecting expression levels of CFTR and other proteins.
https://athenacommons.muw.edu/urc/2026/stem-and-social-sciences-oral-presentations/2