通过RADAR测定隔离和免疫检测酶性DNA-蛋白质交叉链接
Megan Perry1, Gargi Ghosal2,3
1Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE, USA.
Methods in molecular biology (Clifton, N.J.)
|August 13, 2023
概括
研究人员开发了RADAR测试,以检测哺乳动物细胞中的DNA-蛋白交叉链接 (DPC). 这种方法直接量化特定的DPC,帮助研究它们的修复和交叉连接药物的作用.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 生物化学 生物化学
背景情况:
- DNA-蛋白质交叉链接 (DPC) 阻碍了DNA复制和修复.
- 了解DPC的形成和去除对于细胞健康至关重要.
- 现有的检测方法可能缺乏特异性或敏感性.
研究的目的:
- 引入一种用于直接检测和量化哺乳动物细胞中特定DPCs的新型试验.
- 为分析DNA-蛋白质交联活动提供一个敏感的工具.
- 为了促进对DPC修复途径的研究.
主要方法:
- 雷达测试包括从细胞中分离基因组DNA和DPC.
- DNA-DPCs通过真空插槽涂抹在酸纤维素膜上固定不动.
- 检测利用特定于感兴趣的蛋白质的抗体,量化标准化为DNA加载控制.
主要成果:
- 雷达测定可以直接检测特定的酶性DPC.
- 它允许灵敏地分析各种药物诱导的DNA-蛋白质交联活性.
- 该方法可以适应不同的细胞类型和实验条件.
结论:
- 雷达测试是研究DNA-蛋白质交叉链接的宝贵工具.
- 这种技术需要最少的专用设备,提高了可访问性.
- 它促进了对DPC修复机制和药物诱导的交叉连接的理解.
关键词:
修复DNA的修复DNA的修复这是一种DNA-蛋白质交叉链接.DPC维修 DPC维修是什么意思雷达测试方法 雷达测试方法在Slot blot blot中使用.在TOP1-cccc中.在TOP2-cc中.更多相关视频
11:58A Simple, Rapid, and Quantitative Assay to Measure Repair of DNA-protein Crosslinks on Plasmids Transfected into Mammalian Cells
Published on: March 5, 2018
8.3K
10:50Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
Published on: March 9, 2010
17.4K
相关概念视频
02:57Southern Blot
19.6K
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
19.6K
01:33Enzyme-Linked Immunosorbent Assay
13.5K
In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or...
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or...
13.5K