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一种需要少量效应细胞和靶细胞的[35S]甲硫氨酸细胞毒性测定法。

An [35S]methionine cytotoxicity assay requiring small numbers of effector and target cells.

作者信息

Stedman K E, Campbell P A

机构信息

Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, CO 80206.

出版信息

J Immunol Methods. 1989 May 12;119(2):291-4. doi: 10.1016/0022-1759(89)90409-2.

Abstract

This communication describes an assay to detect polyclonal cytotoxic activity of small numbers of effector cells. 200 tumor target cells, radiolabelled with 10,000-20,000 cpm [35S]methionine, are incubated with proportionately few effector lymphocytes in microtiter wells. The targets are sufficiently well labelled to permit detection of cytolysis caused by greater than or equal to 1000 cytotoxic cells, at effector:target ratios as low as 5:1. This method permits the rapid assay of limited numbers of effector cells, avoids lengthy culture of effector cells in order to acquire sufficient numbers, and is useful for limiting dilution analysis of cytotoxic T lymphocytes or their precursors.

摘要

本通讯描述了一种检测少量效应细胞多克隆细胞毒性活性的分析方法。将200个用10,000 - 20,000 cpm [35S]甲硫氨酸进行放射性标记的肿瘤靶细胞,与微量滴定孔中数量相对较少的效应淋巴细胞一起孵育。这些靶细胞标记充分,能够在效应细胞与靶细胞比例低至5:1时,检测出由大于或等于1000个细胞毒性细胞引起的细胞溶解。该方法能够快速检测有限数量的效应细胞,无需为获得足够数量的效应细胞而进行长时间培养,并且对于细胞毒性T淋巴细胞或其前体的有限稀释分析很有用。

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