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Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Updated: Jul 17, 2025

Determining Cell-surface Expression and Endocytic Rate of Proteins in Primary Astrocyte Cultures Using Biotinylation

Determining Cell-surface Expression and Endocytic Rate of Proteins in Primary Astrocyte Cultures Using Biotinylation

Published on: July 3, 2017

细胞表面生物化使用 furan 交联化学.

Esperanza Fernández^1,2, Laia Miret-Casals³, Annemieke Madder³

¹VIB Center for Medical Biotechnology, Ghent, Belgium.

Methods in molecular biology (Clifton, N.J.)

|September 4, 2023

概括

这项研究引入了一种新的表面生物化协议,使用 furan-biotin 亲和力净化. 这种方法增强了血膜蛋白质的丰富性,通过质谱学进行更敏感的表面蛋白质组分析.

关键词:

关联性净化净化方法黄生物是一种这是LC-MS/MS.等离子体膜蛋白质等离子体膜蛋白质蛋白质组学是指蛋白质组学.一些表面的表面.

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Last Updated: Jul 17, 2025

Determining Cell-surface Expression and Endocytic Rate of Proteins in Primary Astrocyte Cultures Using Biotinylation

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科学领域:

蛋白质组学是指蛋白质组学.
细胞生物学细胞生物学
生物化学生物化学

背景情况:

由于蛋白质丰度低和提取困难,细胞表面组分析具有挑战性.
表面蛋白对细胞功能至关重要,但在样本准备过程中经常会丢失.
现有的方法,如表面生物化改善检测,但可以进一步优化.

研究的目的:

开发和验证一种新的,高效的协议,用于丰富血膜蛋白质.
用质谱法提高表面蛋白质组映射的灵敏度和准确性.
为了克服当前表面蛋白质提取和丰富技术的局限性.

主要方法:

开发了一种使用 furan-biotin 亲和度净化方法的新型表面生物化协议.
将该协议应用于可活细胞,用于对表面膜蛋白进行选择性标记.
使用斯特雷普塔维丁珠子丰富生物化蛋白质,然后进行素消化和LC-MS/MS分析.

主要成果:

从细胞表面成功丰富了血膜蛋白质.
与传统方法相比,在绘制表面蛋白质组时表现出更好的灵敏度.
furan-biotin 协议有效地解决了蛋白质聚合和低丰度问题.

结论:

furan-biotin亲和力净化协议为等离子膜蛋白质组学提供了强大的和敏感的方法.
这种方法增强了细胞表面的研究,为细胞表面蛋白质功能提供了更深入的见解.
该协议是推进基于质谱的细胞表面蛋白质蛋白质分析的宝贵工具.