Abstract
Cytosol preparations were made from human intestinal mucosa, and following (NI-I4)2SO4 precipitation were separated by a hollow fibre membrane into a high and a low molecular weight fraction, with a molecular weight separation point at approximately 12,500. The fraction retained by the membrane was further purified using ion exchange and gel chromatography, and the fraction associated with calcium-binding activity further analysed on polyacrylamide gel with and without sodium dodecyl sulphate. Molecular weight determinations on this fraction were performed by gel filtration or by sedimentation equilibrium. (Dr. J. Butler, M.R.C. Laboratory of Molecular Biology, Cambridge). The fraction passing through the membrane was subjected to ion exchange chromatography with and without calcium chloride in the etuting buffer. This step was followed by gel chromatography. Molecular weight was determined by gel filtration and by sodium dodecyl sulphate gel electrophoresis according to Swank and Munkres [4]. Calcium-binding activity was determined by a chelex assay.
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