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Figure 3 Coimmunoprecipitation of Csp and CFTR. Calu-3 membrane extract was mixed with Csp or CFTR antibody and precipitated as described under “Experimental Procedures.” Immunoblots were probed with anti-CFTR (1:2,000, panel A) or anti-Csp antibodies (1:2,000, panel B). Calu-3 cell membrane extract was loaded as a positive control (panels A andB, lanes 1). Antibodies used for immunoprecipitation were nonimmune IgG and anti-Csp (panel A, lanes 2 and 3) and anti-GST and anti-CFTR-NBD1 (panel B, lanes 2 and3). The lower band denoted Csp inpanel B likely represents deacylated Csp, produced by the coimmunoprecipitation conditions, which include overnight incubation with detergent. This outcome was noted by others (43, 47).Hc indicates antibody heavy chain. Image published in: Zhang H et al. (2002) Copyright © 2002. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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