Various types of commercial-grade detergents contain elevated levels of sulfhydryl oxidizing agents, peroxides, salts, and carbonyl compounds. The proteins that are isolated by these detergents are highly susceptible to contaminating peroxides and carbonyls. The peroxides oxidize the proteins, and the carbonyl groups will form Schiff’s bases with the proteins that interfere with its structure.
Topics: Detergents
Blocking buffers are used to bind the surface areas that may attach to reagents during the testing process and prevent nonspecific antibodies from interacting with reagents. Blockers can also cause poor reading or inaccurate results.
In 1975, Porath developed Immobilized Metal Affinity Chromatography (IMAC). It is based on the interaction of specific protein residues, including histidines, cysteines, and occasionally, tryptophans, with cations of transition metals.
Topics: Protein Purification
The RIPA buffer, or Radioimmunoprecipitation assay buffer, is ideal for whole-cell extracts, as well as protein assays. However, it is an essential component in other applications too.
Topics: Protein Extraction
