Preserving Cytonemes for Immunocytochemistry of Cultured Adherent Cells.

Preserving Cytonemes for Immunocytochemistry of Cultured Adherent Cells.
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保存细胞因子用于培养贴壁细胞的免疫细胞化学。

DOI:
10.1007/7651_2020_305
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发表时间:
2021
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Rogers S
Rogers S
中科院分区:
--
文献类型:
--
作者:
Rogers S

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细胞素是一种特殊的信号丝状足,在发育和细胞分化中起作用。然而,它们不能通过标准固定技术很好地保存以研究蛋白质定位和相互作用。最近的一项方法学进展已经产生了改进细胞素保存使用戊二醛固定和硼氢化钠处理,以减少背景。我们在此描述了一种更安全的方法,利用甘氨酸将细胞素在培养的贴壁细胞上戊二醛固定后进行有效阻断,并证明了其在免疫细胞化学中的有效性。
Cytonemes are specialized signalling filopodia that have a role in development and cellular differentiation. However, they are not well preserved by standard fixation techniques to study protein localization and interactions. A recent methodological advance has yielded improvements in cytoneme preservation using glutaraldehyde fixation and sodium borohydride treatment to reduce background. We herein describe a safer method for effective blocking using glycine following glutaraldehyde fixation of cytonemes on cultured adherent cells and demonstrate its effectiveness in immunocytochemistry.
发育中的细胞因子
DOI: --
发表时间: 2019
影响因子: 4
作者:
Chengting Zhang;S. Scholpp
通讯作者: S. Scholpp