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标记微孔板中的细胞以测定[3H]胸腺嘧啶核苷摄取量。

Labeling cells in microtiter plates for determination of [3H]thymidine uptake.

作者信息

Shevach E M

机构信息

National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.

出版信息

Curr Protoc Immunol. 2001 May;Appendix 3:Appendix 3D. doi: 10.1002/0471142735.ima03ds21.

Abstract

A number of protocols in Current Protocols in Immunology use as their end-point the determination of cell proliferation by determining the incorporation of [(3)H]thymidine into cellular DNA. This appendix presents a protocol in which the radioactive label is added during the last 4 to 24 hr of the culture. A semiautomated cell harvesting apparatus is then used to lyse the cells with water and precipitate the labeled DNA on glass fiber filters. The filter pads are then dried and counted by standard liquid scintillation counting techniques in a scintillation counter.

摘要

《免疫学实验指南》中的许多实验方案都以通过测定[³H]胸腺嘧啶核苷掺入细胞DNA来确定细胞增殖作为终点。本附录介绍了一种在培养的最后4至24小时添加放射性标记的实验方案。然后使用半自动细胞收获装置用水裂解细胞,并将标记的DNA沉淀在玻璃纤维滤膜上。然后将滤膜垫干燥,并通过闪烁计数器中的标准液体闪烁计数技术进行计数。

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