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利用流式细胞术根据细胞群体中硫酸乙酰肝素表位的表达情况对其进行表征和分离。

Use of flow cytometry for characterization and fractionation of cell populations based on their expression of heparan sulfate epitopes.

作者信息

Holley Rebecca J, Smith Raymond A, van de Westerlo Els M A, Pickford Claire E, Merry C L R, van Kuppevelt Toin H

机构信息

Wellcome Trust Centre for Cell Matrix Research, Faculty of Life Sciences, University of Manchester, Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK.

出版信息

Methods Mol Biol. 2015;1229:239-51. doi: 10.1007/978-1-4939-1714-3_21.

Abstract

The ability to characterize alterations in heparan sulfate (HS) structure during development or as a result of loss or mutation of one or more components of the HS biosynthetic pathway is essential for broad understanding of the effects these changes may have on cell/tissue function. The use of anti-HS antibodies provides an opportunity to study HS chain composition in situ, with a multitude of different antibodies having been generated that recognize subtle differences in HS patterning, with the number and positioning of sulfate groups influencing antibody binding affinity. Flow cytometry is a valuable technique to enable the rapid characterization of the changes in HS-specific antibody binding in situ, allowing multiple cell types to be directly compared. Additionally fluorescent-activated cell sorting (FACS) allows fractionation of cells based on their HS-epitope expression.

摘要

在发育过程中或由于硫酸乙酰肝素(HS)生物合成途径中一种或多种成分的缺失或突变而能够表征HS结构的改变,对于广泛理解这些变化可能对细胞/组织功能产生的影响至关重要。使用抗HS抗体提供了一个原位研究HS链组成的机会,已经产生了多种不同的抗体,它们能够识别HS模式中的细微差异,硫酸基团的数量和位置会影响抗体的结合亲和力。流式细胞术是一种有价值的技术,能够快速表征原位HS特异性抗体结合的变化,允许直接比较多种细胞类型。此外,荧光激活细胞分选(FACS)允许根据细胞的HS表位表达对细胞进行分离。

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