Cytoplasmic Protein Extraction

Cytoplasma is the cell interior place between the cellular membrane and the nucleus, where various intracellular activity take place. Therefore, cytoplasmic protein extraction is important to study biochemical reactions inside cells, such as energy metabolism, heme-synthesis, nitrogen and lipid metabolism, phospholipids biosynthesis, and other various protein-protein interactions. In pharmaceutical industry, this procedure has enormous merits for drug design with potential targets identified and characterized.

A robust technique to separate highly soluble (hydrophilic) cytoplasmic proteins from poorly soluble (hydrophobic) membrane-associated proteins is to use differential solubility in a progressive series of extraction buffers, each containing more potent solubilizing chaotropes and detergents. It is known as the "sequential extraction" procedure. After cell lysis, solubilization agents such as Triton X-100 are sequentially added and centrifugated to separate cytoplasmatic fractions and nuclei by dissolving membrane-associated proteins. After that, cytoplasmatic proteins can be precipitated from the hydrophilic fraction by chemicals such as trichloroacetic acid (TCA). For isolation of specific proteins, such as metalloproteomics, further steps need to be designed based on the properties of targets. Gel-based or gel-free technology, mass spectrometry as a good example, is applied for analysis and characterization after extraction.

Creative BioMart is specialized in proteomic analysis, including cytoplasmic proteomics and related cytoplasmic protein extraction. Having years of experience with different organisms and cell lines, we provide cytoplasmic proteins isolation and high-quality analysis. We also provide expression, purification and structure analysis service for specific proteins. Creative BioMart offers professional one-stop services for our customer:

• Cytoplasmic protein isolation.
• Proteomic analysis with gel-based or gel-free technology.
• Protein identification and characterization.
• Ready-to-use cytoplasmic proteins from various organisms.
• Expression, purification and structure analysis.

Reference:

1. Cordwell S.J. (2008) Sequential Extraction of Proteins by Chemical Reagents. In: Posch A. (eds) 2D PAGE: Sample Preparation and Fractionation. Methods in Molecular Biology™, vol 424. Humana Press.