Common Types of Proteins Used in WB Experimental Protein Extraction
When performing protein extraction for WB experiments, the commonly extracted protein fractions include the following:
Total Cellular Proteins
This is the most commonly extracted protein fraction, obtained by lysing both the plasma and nuclear membranes to release all intracellular proteins. This extraction approach is widely used to analyze overall protein expression in cells.
Nuclear Proteins
Nuclear proteins are isolated by selectively lysing the cytoplasmic membrane while preserving nuclear integrity, followed by centrifugation to separate nuclear components. This extraction method is particularly suitable for studying proteins involved in nuclear processes, such as transcription factors and DNA repair proteins.
Membrane Proteins
The cell membrane serves as a critical interface between the intracellular and extracellular environments, hosting proteins essential for signaling, transport, and structural integrity. The isolation of membrane proteins requires specialized extraction techniques, such as detergent-based solubilization or phase separation methods, to effectively enrich these proteins.
Organelle-Specific Proteins
Each organelle within a cell has distinct functions and contains a unique set of proteins. By employing differential centrifugation or gradient-based separation, organelles such as mitochondria, the endoplasmic reticulum, and the Golgi apparatus can be isolated, followed by targeted protein extraction.
Extracellular Proteins
Certain proteins are secreted into the extracellular space, including cytokines, growth factors, and extracellular matrix proteins. The extraction of extracellular proteins involves processing culture supernatants or body fluids to concentrate and purify these proteins for further analysis.
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