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.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
