Malik N, Bustin M, Smulson M
Nucleic Acids Res. 1982 May 11;10(9):2939-50. doi: 10.1093/nar/10.9.2939.
To facilitate investigations on the organization of poly (ADP-Rib) polymerase in chromatin, and to elucidate its biological function, polymerase purified from HeLa nuclei was used to elicit antibodies in mice. The anti-polymerase sera was found to be specific by multiple criteria. The association of polymerase with oligonucleosomes of differing chain size was determined by the specific binding of polymerase antibody (and as control, anti-histone H3) to nitrocellulose transfers of native electrophoretic gels of these particles. By this technique, polymerase appears to be associated with a select subclass of nucleosomes. Further resolution of the polymerase binding sites within nucleosome classes was achieved with the antibody by two-dimensional native polyacrylamide gel electrophoresis and nitrocellulose transfer.
为便于研究聚(ADP - 核糖)聚合酶在染色质中的组织方式,并阐明其生物学功能,从HeLa细胞核中纯化的聚合酶被用于在小鼠体内诱导产生抗体。通过多种标准发现抗聚合酶血清具有特异性。通过聚合酶抗体(以及作为对照的抗组蛋白H3)与这些颗粒的天然电泳凝胶在硝酸纤维素膜上的转移产物的特异性结合,确定了聚合酶与不同链大小的寡核小体的关联。通过这种技术,聚合酶似乎与核小体的一个特定亚类相关联。利用该抗体通过二维天然聚丙烯酰胺凝胶电泳和硝酸纤维素膜转移,进一步解析了核小体类别内的聚合酶结合位点。