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Cystic Fibrosis: Diagnosis and Protocols, Volume I: by Kris De Boeck, Melissa Ashlock (auth.), Margarida D. Amaral,

By Kris De Boeck, Melissa Ashlock (auth.), Margarida D. Amaral, Karl Kunzelmann (eds.)

Despite the numerous milestones in cystic fibrosis (CF) examine, development in the direction of curing the sickness has been sluggish, and it really is more and more tough to understand and use the already broad and nonetheless starting to be diversity of numerous tools at present hired to check CF so one can know it in its multidisciplinary nature. Cystic Fibrosis: prognosis and Protocols goals to supply the CF study neighborhood and comparable researchers with a truly wide variety of top quality experimental instruments, as an ideal way to know and use classical and novel equipment utilized to cystic fibrosis. Volume I: ways to check and proper CFTR Defects makes a speciality of the cystic fibrosis transmembrane conductance regulator (CFTR) and its expression, biogenesis, constitution, and serve as by way of the defects inflicting CF. Written within the hugely winning Methods in Molecular Biology™ sequence layout, chapters comprise introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, simply reproducible laboratory protocols, and pointers on troubleshooting and fending off identified pitfalls.

Comprehensive and functional, Cystic Fibrosis: analysis and Protocols will offer readers with optimum operating instruments to deal with urgent questions within the top technical method, whereas supporting we all, as a examine and medical neighborhood, to maneuver quicker hand-in-hand towards unravelling the secrets and techniques of this not easy illness and medication it.

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Additional info for Cystic Fibrosis: Diagnosis and Protocols, Volume I: Approaches to Study and Correct CFTR Defects

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4 U/ml). 3. AccutaseTM (Innovative Cell Technologies, Catalog # AT104).

The precipitate will reduce the transfection efficiency, so it should be removed by filtering the media used for transfection. 4. Expression is best if the cells are actively growing and very poor if the cells are 100% confluent. 5. , COS-1 cells) because they do not attach very well to the bottom of the flask. Therefore, one has to be gentle when moving the cells from the incubator to the culture hood. An advantage is that trypsin is not required to dislodge the cells from the bottom of the flask.

Cell incubation and assay are carried out at 37◦ C. 4. CFTR Assay by Fluorescence Microscope This method has been successfully used to study the effect of CFTR inhibitors (10) and potentiators (11) after transient transfection in cells like HEK-293: 1. At the time of assay (48 h after transfection in 96-well microplates), cells are washed twice with standard Dulbecco’s PBS. 2. , forskolin, CPT-cAMP, and potentiators). 3. After this step, the microplates carrying the cells are transferred to the microscope to perform the assay, one well at a time.

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