Isolation of Aged Yeast Cells Using Biotin-Streptavidin Affinity Purification.

Isolation of Aged Yeast Cells Using Biotin-Streptavidin Affinity Purification.
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使用生物素-链霉亲和素亲和纯化分离老化酵母细胞。

DOI:
10.1007/978-1-0716-0868-5_17
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发表时间:
2021
期刊:
Yeast Protocols. Methods in molecular biology
影响因子:
--
通讯作者:
Beidong Liu
Beidong Liu
中科院分区:
其他
文献类型:
--
作者:
Xuejiao Jin;Xiuling Cao;Beidong Liu

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酵母(酿酒酵母)已被用作研究细胞衰老分子机制的主要模型系统之一。研究酵母衰老的一个主要技术挑战是从指数生长的细胞培养物中分离衰老细胞,因为这种培养物中的衰老细胞很少见。为了实现这一目标,已经开发了几种分离老化细胞的方法。在这里,我们描述了用于分离老化酵母细胞的生物素-链霉亲和素亲和纯化方案。它由三个主要步骤组成:酵母细胞的生物素化、将细胞培养至所需年龄以及使用链霉亲和素包被的磁微珠收获老化细胞。分离的老化细胞可用于显微镜、生物化学或分子生物学分析。
Yeast (Saccharomyces cerevisiae) has been used as one of the main model systems for studying molecular mechanisms underlying cellular aging. A major technical challenge in studying aging in yeast is the isolation of aged cells from exponentially growing cell cultures, since aged cells in such cultures are rare. Several methods for isolating aged cells have been developed to achieve this. Here, we describe a biotin-streptavidin affinity purification protocol for isolating aged yeast cells. It consists of three main steps: biotinylation of yeast cells, culturing cells to the desired age, and harvesting the aged cells using streptavidin-coated magnetic microbeads. The isolated aged cells can be used for microscopy, biochemistry, or molecular biology analysis.
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发表时间: 2012-07-03
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