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. Author manuscript; available in PMC: 2014 Jul 9.
Published in final edited form as: J Biol Chem. 2008 Dec 4;284(6):3640–3649. doi: 10.1074/jbc.M806580200

Figure 1. Expression and ATPase activity of ABCA4 and ABCA4-1D4.

Figure 1

ABCA4 and ABCA4-1D4 containing a 9 amino acid 1D4 epitope were expressed in HEK 293T cells, solubilized in CHAPS detergent, purified on a Rim 3F4 – Sepharose immunoaffinity matrix and reconstituted into liposomes for determination of basal and all-trans retinal stimulated ATPase activity measurements. A. SDS gel stained with Coomassie blue showing a typical membrane extract from HEK 293T cells expressing ABCA4 (lane 1), immunoaffinity purified ABCA4 (lane 2), and immunoaffinity purified ABCA4-1D4 (lane 3). B. Relative expression levels of ABCA4 and ABCA4-1D4 quantified on Western blots of membranes from HEK293 cells expressing these proteins. C. ATPase activity of purified and reconstituted ABCA4 and ABCA4-1D4 as a function of added all-trans retinal. For each assay 20–40 ng of protein was used. Data show the average value for 3 experiments ± SD.