Purification and N-terminal amino acid sequence of proliferating cell nuclear antigen (PCNA)/cyclin and development of ELISA for anti-PCNA antibodies.

摘要: Proliferating cell nuclear antigen (PCNA), also called cyclin, was purified from PBS extract of rabbit thymus by using a combination ammonium sulfate fractionation, DEAE-Sephacel, HPLC ion exchange, and gel filtration column chromatography. PCNA more than 600 times analyzed sodium dodecyl polyacrylamide electrophoresis (SDS-PAGE) immunoblotting. SDS-PAGE showed that 36 kD protein selectively isolated in this purification process, identified as Other previously antigens, Sm, nRNP, SS-A/Ro, SS-B/La, histone, DNA, were not detected preparation counterimmunoelectrophoresis enzyme-linked immunosorbent assay (ELISA). Purified used an to develop ELISA for rapid specific detection anti-PCNA human sera. For further purification, the band electrophoretically eluted SDS slices. The amino acid composition first 25 residues N-terminus determined electroeluted PCNA. This sequence found be unique little homology with existent proteins identification resources databank.