ANALYTIC EXTRACTOR KIT
Size : 1KT
Brand : Anatrace
Integral membrane proteins (IMPs) insert into biological membranes such that the cell membrane, which is a lipid bilayer approximately 30 Å in thickness, shields the hydrophobic membrane-spanning regions of the protein. To purify IMPs, the protein must first be extracted from the bilayer through the replacement of cell lipids with solubilizing detergents. The selection of detergent for both the initial solubilization of the IMP and for downstream study is exceptionally important, as one must ensure that the selected detergent does not inhibit function, cause irreversible denaturation or interfere with protein purification or crystallization.
Previous studies on the selection of detergents for the initial extraction of IMPs have demonstrated the importance of detergent screening (REFS). One of the most exhaustive studies was performed by White et Al. (2007) in which 122 high-expressing predicted membrane proteins were screened in 6 commonly used detergents Triton X-100, LDAO, FC-12, C8E4 and DDM. In these tests, the zwitterionic detergetns LDAO and FC-12 were the most efficient at solubilizing yeast membrane proteins; however, all of the detergents in the panel solubilized at least 35% of the proteins tested.
The Analytic Extractor kit contains a panel of eight detergents commonly used for the solubilization of IMPs from the cell membrane. The detergents in this panel were chosen based on their prior effectiveness and use in the literature, and the working concentration of each detergent is dependent on the critical micelle concentration (CMC) of each detergent (i.e. higher concentrations of lower CMC detergents compared to higher CMC detergents) (Ref to Wiener, 2004). The results from this assay will aid in the selection of a detergent for further downstream studies, including x-ray crystallography, NMR, and biochemical assays.