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相关概念视频

Labeling DNA Probes03:31

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...
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.

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相关实验视频

Updated: Jun 16, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
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优化多功能光配体用于细胞内标签.

Pratik Kumar1, Jason D Vevea2,3,4, Ariana N Tkachuk1

  • 1Janelia Research Campus, HHMI, Ashburn, VA 20147.

Proceedings of the National Academy of Sciences of the United States of America
|October 27, 2025
PubMed
概括

研究人员开发了细胞透的光标签,用于标记活细胞中的蛋白质. 这些新工具允许蛋白质净化和有针对性的核转移,增强细胞研究能力.

关键词:
这是HaloTag的标签.化学生物学 化学生物学化学 化学 化学 化学光是一种光.蛋白质净化 蛋白质净化

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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

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Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging
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Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging

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相关实验视频

Last Updated: Jun 16, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

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Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging
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Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging

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科学领域:

  • 化学生物学 化学生物学
  • 分子生物学分子生物学
  • 药用化学 医学化学

背景情况:

  • 基于酶的自我标签标签有助于合成分子对活细胞内的蛋白质进行共价附着.
  • 设计具有光体,亲和度标签或药理学剂的细胞透多功能连接物是具有挑战性的,因为潜在的膜透性问题.

研究的目的:

  • 为了研究基于罗达胺的自我标签标签配体的化学特性,以改善细胞透性.
  • 设计和验证细胞透的多功能连接物用于细胞内蛋白质操纵.

主要方法:

  • 罗达胺基联体的药物化学分析,重点关注乳-特平衡常数 (K_L-Z) 和分布系数 (logD_7.4).
  • 设计和合成包含生物素或JQ1部分的多功能HaloTag配体.
  • 细胞实验以评估联结体介导的蛋白质转位和转录活性.

主要成果:

  • 在K_L-Z和logD_7.4之间观察到负相关性,表明低K_L-Z和高logD_7.4染料 (例如Si-rhodamines) 增强细胞进入.
  • 成功设计了细胞透的HaloTag配体,用于线粒体净化 (生物素) 和BRD4 (JQ1) 的核转位.
  • 将BRD4转移到构成性异色染色体导致转录活性增加.

结论:

  • 该研究提供了设计原则,用于创建细胞透的多功能光联体.
  • 这些新型试剂能够捕获亲和力,并在活细胞中向转移细胞内蛋白质.
  • 开发的化学工具将通过精确操纵细胞组件来推动生物研究.