A computer program for the calculation of sedimentation coefficients and molecular weights of nucleic acids |
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Authors: | B D Young R Krumlauf |
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Affiliation: | Department of Cellular Pathology, Imperial Cancer Research Fund Laboratories, Lincoln''s Inn Fields, London, WC2 3PX, United Kingdom, England |
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Abstract: | A procedure is described for the purification of nuclei and identification of chromatin proteins in transformed epithelial cell lines from mammalian bladder and salivary gland. Nuclear purification was performed by homogenization, in hypotonic buffer containing polyamines to stabilize the nuclear structure, followed by 0.1–0.2% Triton X-100 washing and centrifugation through 2.2 m sucrose. Chromatin was liberated from nuclei by freeze-thawing in hypotonic buffer and the chromatin proteins were extracted with 7 m urea/3 m NaCl. The chromatin proteins were identified using NEPHGE two-dimensional electrophoresis and fluorographic autoradiography. This procedure enabled detection of histones and a range of basic nonhistone chromatin proteins, following cell culture in the presence of low levels of l-(4,5-3H)leucine. |
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Keywords: | Current address and address for reprints: Queen Elizabeth II Fellow School of Pathology University of New South Wales P.O. Box 1 Kensington New South Wales 2033 Australia. |
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