1985 Volume 29 Issue 1 Pages 75-81
The U1-snRNP particles from the nuclear extracts of human KB cells and mouse teratocarcinoma embryoid bodies (OTT6050), purified by SLE-anti-(U1)RNP antibody affinity chromatography, were analyzed for their protein components. The purified human U1-snRNP was recovered from the column by elution with 2.5M MgCl2 at pH7.2 after the column had been washed with Tris-buffer containing DOC, Triton X-100 and 1.0M NaCl. The 3H amino acid-labeled protein components of human (KB) U1-snRNP resolved into three major bands with apparent molecular weights of 69, 000, 58, 000 and 35, 000 upon SDS-polyacrylamide gel electrophoresis.
Highly purified U1-snRNP particles containing U1a and U1b RNAs as their sole nucleic acid moieties were obtained from the mouse teratocarcinoma embryoid bodies by the same procedure as used for purification of KB cell U1-snRNP. By gel electrophoresis, five major polypeptides, with apparent molecular weights of 80, 000, 76, 000, 72, 000 58, 000 and 39, 000, were detected associated with U1-snRNP(s) of embryoid bodies.
