| Autor: |
Zhaodi Jiang, Li-Lin Du, Xiangbing Qi, Gaihong Cai, Pei Zhao, Ying Liu, Ying-Ying Wang, Xiaobin Sun, Xiao-Man Liu, Yan Liu, Xiumei Jin, Xinbin Cai, Ming Li, Yanyong Hu, Yuhua Li, Yaqi Tang, She Chen, Sitong Liu, Wanzhong He |
| Rok vydání: |
2020 |
| Předmět: |
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| Popis: |
We developed a novel auto-nucleation suppressed mechanism (ANSM) for direct synthesis of EM-visible gold nanoparticles (AuNPs) on cysteine-rich tags (e.g., metallothionein) in cells for single-molecule detection with electron microscopy (it accompanies our Nature Method manuscript, Jiang et al. 2020 [1]). Both tagged-fusion proteins expressed in cells (e.g.bacteria, yeast and mammalian cells) and antigens stained with antibody-tag fusion proteins can be visualized by this protocol. Here we describe the typical protocols (both the chemical fixation and the high pressure freezing cases) developed for ANSM-based AuNP synthesis in yeast cells expressing metallothionein (MTn) tags (Figure 1). This approach should be useful for EM visualization of single-molecule in yeast cells, and easier adapted for bacterial cells. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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