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相关概念视频

Subcellular Fractionation01:32

Subcellular Fractionation

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The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...
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相关实验视频

Updated: May 4, 2026

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
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细胞内体分析的方法

Rosangela A S Eichler1, Luiz Felipe Martucci1, Leandro M de Castro2

  • 1Department of Pharmacology, Biomedical Sciences Institute, University of São Paulo, São Paulo, SP, Brazil.

Methods in molecular biology (Clifton, N.J.)
|March 29, 2024
PubMed
概括
此摘要是机器生成的。

提取细胞内需要快速失活降解酶. 将样品加热到80°C对大多数组织起作用,但血需要其他方法来防止凝血和保持的完整性.

关键词:
脑子 脑子 脑子 脑子甲甲是一种形式的甲.细胞内类.血等离子体的使用方法它是稳定的同位素.

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Semi-Quantitative Analysis of Peptidoglycan by Liquid Chromatography Mass Spectrometry and Bioinformatics
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科学领域:

  • 生物化学 生物化学
  • 分子生物学分子生物学
  • 蛋白质组学是指蛋白质组学.

背景情况:

  • 细胞内是从蛋白质降解中产生的生物活性分子.
  • 这些的高效提取对于它们的研究至关重要.
  • 生物样本中的蛋白酶和酶可以快速降解.

研究的目的:

  • 描述提取细胞内的有效方法.
  • 为了应对在提取过程中降解的酶失活的挑战.

主要方法:

  • 蛋白酶和酶的快速失活是必不可少的.
  • 将生物源 (组织,细胞,液体) 在80°C温度下加热20分钟有效地使酶变质.
  • 避免在高温下酸化,可以防止非特异性水解,特别是Asp-Pro键的水解.

主要成果:

  • 对于大多数生物样本来说,在80°C的热失活是足够的.
  • 由于高于55°C的热引起的凝血,等离子体样本需要使用其他酶非激活方法.
  • 详细介绍了成功提取细胞内的具体协议.

结论:

  • 有效的细胞内提取依赖于快速的酶失活.
  • 样品类型决定了酶失活的适当方法.
  • 描述的方法有助于恢复生物活性细胞内.