Novel cell lysis buffer solution for detecting cell ubiquitin modified proteins

Abstract

The invention discloses a novel cell lysis buffer solution for detecting cell ubiquitin-like modified proteins, a large number of micromolecule modified proteins, especially ubiquitin-like molecule modified proteins exist in human and animal cells, and in order to improve the detection efficiency, the composition proportion of each component of the animal cell lysis buffer solution is optimized and adjusted, related reagent components are creatively added, the cell lysis process is changed and improved, various animal primary generation cells and subculture cells can be highly and effectively split by using the cell lysis method, the related reagent components are innovatively added, and the cell lysis process is changed and perfected, such that the good experimental early-stage protein sample is provided for the later-stage experiments such as the protein identification, the protein purification, the antibody preparation and the like.

Description

technical field [0001] The invention belongs to the technical field of cell detection, and in particular relates to a novel cell lysis buffer for detecting cell ubiquitin-modified proteins. Background technique [0002] There are a variety of ubiquitin-like small protein molecules in cells, including Ubiquitin, SUMO, ISG, etc., which are covalently linked to substrate protein molecules through cascade reactions in cells, thereby regulating the molecular functions of substrate proteins. Therefore, accurate and efficient identification of protein substrates modified by ubiquitin molecules is of great significance in basic biomedical research and clinical practice. [0003] The technical methods commonly used to identify proteins modified by ubiquitin-like molecules in cells mainly include immunoblotting, co-immunoprecipitation, and protein purification. However, the implementation of all these experimental techniques is based on the acquisition of suitable cellular protein sa...

AI Technical Summary

Problems solved by technology

However, because the protein modified by ubiquitin molecules in the cell only accounts for 0.1-5% of the total protein, and is usually in a dynamic balance between modification and de-modification; at the same time, due to the presence of a large number of Deubiquitin-modifying enzymes, these enzymes can cause deubiquitination of ubiquitin-modified proteins during cell lysis, resulting in the use of these cell lysis buffers to obtain cellular protein samples in most cases only Can detect proteins that are not modified by ubiquitin-like molecules, but not proteins that are modified by ubiquitin-like molecules

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