Holley Rebecca J, Smith Raymond A, van de Westerlo Els M A, Pickford Claire E, Merry C L R, van Kuppevelt Toin H
Wellcome Trust Centre for Cell Matrix Research, Faculty of Life Sciences, University of Manchester, Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK.
Methods Mol Biol. 2015;1229:239-51. doi: 10.1007/978-1-4939-1714-3_21.
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.
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。
