PMC:13922 / 18083-18922 JSONTXT

{"project":"Colil","denotations":[{"id":"T25","span":{"begin":306,"end":308},"obj":"5641522"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/docs/sourcedb/PubMed/sourceid/"}],"text":"SDS-PAGE analysis for tissue oestrogen receptors\nSliced breast cancer tissues (thickness varied among samples, but was always less than 0.5 mm) were incubated for 1 h at 37°C with [125I]TAZ (final concentration 1 nmol/l) in Krebs-Ringer phosphate buffer (pH 7.4), with 2% bovine albumin and 0.25% glucose [31,32]. Then, all tissues were washed with Krebs-Ringer phosphate buffer and mixed with the same solution, but containing 1% SDS, 1.6 mmol/l EDTA and 2% β-mercaptoethanol.Tissue slices were briefly and delicately homogenized, and heated to 100°C for 2 min. Protein extraction was carried out by phenol and precipitation by acetone containing 0.1 mol/l acetic acid [32]. After 2 h at -20°C, the extracts were centrifuged. After washing, the pellets were solubilized in lysis buffer and subjected to electrophoresis as described above."}