通过免疫沉,然后通过质谱测量来识别蛋白质素化协议
Alexandra Indeglia1, Hsin-Yao Tang2
1Program in Molecular and Cellular Oncogenesis, The Wistar Institute, Philadelphia, PA 19104, USA; Graduate Group in Biochemistry and Molecular Biophysics, the University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
STAR protocols
|January 13, 2026
概括
这项研究详细介绍了一种识别蛋白质素化方法,这种方法是由基氨酸脱胺酶 (PAD) 酶所发生的修饰. 该协议使用免疫沉和质谱测量来确定蛋白质中的素残留物.
科学领域:
- 生物化学 生物化学
- 蛋白质组学是指蛋白质组学.
- 分子生物学分子生物学
背景情况:
- 蛋白质素化是一种翻译后的修改,将氨酸转化为氨酸.
- 这一过程是由基氨酸脱敏酶 (PAD) 酶催化.
- 氨酸在各种生理和病理过程中起作用.
研究的目的:
- 建立一个强大的协议,用于识别素蛋白残留物.
- 为了能够在体外和细胞培养中检测出素修饰的变化.
- 提供一种免疫沉和基于质谱的素蛋白质鉴定方法.
主要方法:
- 氨酸蛋白质的免疫沉.
- 液体染色学-双重质谱学 (LC-MS/MS) 用于残留物识别.
- 该协议应用于重组蛋白和细胞培养试验.
主要成果:
- 通过使用描述的协议,成功识别了素蛋白残留物.
- 在体外和细胞培养环境中证明该协议的有效性.
- 免疫沉和质谱分析的详细步骤.
结论:
- 本协议有效地识别了素蛋白残留物.
- 该方法适用于各种实验系统,包括重组蛋白和细胞培养.
- 该协议有助于研究基氨酸减弱酶活性及其后果.
更多相关视频
05:43A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis
Published on: January 24, 2017
8.9K
11:54Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
Published on: March 23, 2020
10.2K
相关概念视频
Enzyme-Linked Immunosorbent Assay
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 quantified.
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 quantified.
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.
