Sample Results

As a test of the impedance hardware and software we prepared an analog circuit with resistors and capacitors with known measured values. This circuit was connected to the equipment and an impedance spectrum was obtained. The impedance values were then fitted using BLIMP to obtain estimates of the resistances and capacitances. As shown in Fig. 1, the equipment and software gave excellent estimates of the known values and an excellent fit (solid line) of the impedance spectrum. From this Nyquist...

Generation of Mice of the Appropriate Genotype

The objective of the breeding program is to generate second generation pups that carry at least one copy of the SV40 transgene and are either non-CF (CFTR + +, or + -) or CF (CFTR - -). A male ImmortoMouse is placed with three female mice heterzygous for the S489X CFTR mutation in a microisolator cage. Multiple breeding cages are established to increase the number of first-generation offspring. When the female mice become pregnant, they are transferred to individual cages and are replaced with...

Cell Culture and Expansion

Ca2+- and Mg2+-free Hanks balanced salt solution that contains 0.05 trypsin and 0.53 mM EDTA (Gibco-BRL). 3. Collecting tubule culture media. 1 1 mix of Dulbecco's modified Eagle's medium (DMEM) and Ham's F-12 medium (GIBCO-BRL) supplemented with 1.3 yg L sodium selenite, 1.3 yg L 3,3',5-triiodo-L-thyronine, 5 mg L insulin, 5 mg L transferrin, 25 yg L prostaglandin E1, 2.5 mM glutamine, 5 nM dexamethasone, 50000 U L nystatin, 50 mg L streptomycin, 30 mg L...

Reconstitution of CFTR Protein into Liposomes from PFO

To study the structure and function of a purified membrane protein, it is essential that the detergent in which the protein is purified be exchanged for phospholipids. This exchange is a time- and concentration-dependent process. Typically, phospholipids are added to detergent-solubilized proteins and then the detergent is removed by dialysis or detergent adsorption resins 24,25 . Only when the detergent concentration decreases below a critical value critical micelle concentration or CMC can...