在体外和体内使用编码的标签进行蛋白质的光标签
Maya de Luis1, Shuwa Xu1, Kai Zinn1
1Division of Biology and Biological Engineering, California Institute of Technology.
The Journal of biological chemistry
|May 16, 2025
概括
皮质标签标记蛋白质用于识别和净化. 本综述探讨了使用光显微镜在固定和实时成像中进行蛋白质动力学研究的抗体,纳米体和小分子标签.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 显微镜的使用方法
背景情况:
- 皮标签是用于蛋白质标签和操纵的多功能工具.
- 它们在各种生物环境中促进了蛋白质的识别,净化和可视化.
- 传统的标签往往缺乏研究活细胞中的蛋白质动态的能力.
研究的目的:
- 审查光显微镜中使用的表位标签.
- 讨论对抗体,纳米体和小分子有亲和力的标签.
- 突出它们在固定细胞和活细胞成像中的应用,以研究蛋白质动力学.
主要方法:
- 在光显微镜中的表位标签应用的文献综述.
- 讨论不同的标签类型及其绑定亲和关系.
- 分析它们在固定细胞和活细胞成像技术中的有用性.
主要成果:
- 位标签通过光显微镜使蛋白质定位和动力学研究成为可能.
- 与抗体结合的标签对于固定样本免疫组织化学有用.
- 设计用于实时成像的标签提供了对蛋白质动态的洞察,这在固定的样本方法中是不可能的.
结论:
- 位标签对于可视化蛋白质定位和动态至关重要.
- 标签的选择取决于是否需要固定或实时成像.
- 先进的表位标签为实时研究蛋白质行为提供了新的可能性.
相关概念视频
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...
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...
Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
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...
Protein Dynamics in Living Cells
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...


