Protein extraction is an important step in any proteomics experiment. It often starts with cell lysis and cell fractionation, followed by specific enrichment and/or isolation of a particular protein of interest (affinity purification), or removal of interfering or contaminating substances (i.e. immunodepletion).
Many techniques are available for the disruption of cells, including
physical and detergent-based methods. Historically, physical lysis has been
the method of choice for cell disruption; however, physical methods often
require expensive equipment (french prss, homogenizer, cryogrinding).
Detergent-based lysis is more popular due to ease of use, low cost and efficient protocols. However, many detergents interfere with the downstream LC-MS analysis. A list of MS compatible detergents can be found on our protein digestion page.
Overviews by Life Technologies (former Pierce, now Thermo)
Sample Preparation for Mass Spectrometry page
Mass Spectrometry Sample Preparation Handbook page
Physical cell lysis page
Detergent based cell lysis page
Protein Extraction from Tissues and Plants
Cell culture related protocols
Protein preparation from Body Fluids
blood, plasma, urine, CSF etc
Isotopic labelling strategies
SILAC (stable isotope labeling using amino acids in cell culture) is a metabolic labelling technique for comprehensive identification, characterization and quantification of proteins by LC-MS/MS. Isotopically labeled amino acids (typically Lys and Arg) are incorporated in to proteins during cell culture.