兔和鸡雌激素受体及相关肽的多克隆抗体。用于雌激素受体的亲和分离和与雌激素受体相关的染色质片段的检索。

Receptor Pub Date : 1992-01-01
T J Schuh, H Ahrens, M D Mogensen, J Gorski, G C Mueller
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引用次数: 0

摘要

从鸡和兔制备了针对人雌激素受体(hER)两个区域多肽的多克隆抗体。作为抗原的雌激素受体(ER)肽在大肠杆菌中过量产生。如有需要,抗体是用含有细菌包涵体抗原的树脂高产量偶联到环氧活化琼脂糖上的亲和纯化的。抗体在免疫印迹实验中识别变性人、牛、大鼠和兔内质网。兔抗血清和固定蛋白a很容易实现天然内质网蛋白的免疫沉淀。鸡抗体,可大量获得,偶联琼脂糖后也可用于免疫分离。通过使用这些试剂,已经实现了从MCF-7细胞中选择性检索与内质网相互作用的染色质片段。
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Polyclonal antibodies from rabbits and chickens against the estrogen receptor and related peptides. Use in the affinity isolation of estrogen receptors and the retrieval of chromatin fragments associating with estrogen receptors.

Polyclonal antibodies from chickens and rabbits have been prepared against polypeptides representing two regions of the human estrogen receptor (hER). The estrogen receptor (ER) peptides used as antigens were overproduced in Escherichia coli. When indicated, the antibodies were affinity purified using resins to which the antigens contained in bacterial inclusion bodies had been coupled in high yield to epoxy-activated agarose. The antibodies recognize denatured human, bovine, rat, and rabbit ER in immunoblotting experiments. Immuno-precipitation of native ER protein was readily accomplished using rabbit antisera and immobilized protein A. The chicken antibodies, available in larger quantities, were also useful for immunoisolation after coupling to agarose. With the use of these reagents, the selective retrieval of chromatin fragments from MCF-7 cells that interact with ER has been achieved.

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