Purification of membrane proteins.
As stated at the beginning of this chapter, it is not possible to present a single step-by-step protocol for the purification of all membrane proteins. We have discussed many of the individual techniques employed to purify membrane proteins and the problems associated with their application. A successful purification protocol will require the use of a combination of these techniques. It should be clear from the examples given, however, that affinity chromatography is by far the most useful technique available. The greatest single obstacle to performing a successful purification is the ability to maintain solubilized proteins in fully dispersed monomeric micelles without inactivating the protein. The best combination of techniques and the optimal conditions for their use can be determined only by trial and error. This process should become easier as a greater number of examples become available and as greater use is made of HPLC and FPLC techniques.
Thomas
T C
TC
McNamee
M G
MG
D002458:Cell Fractionation / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D007158:Immunologic Techniques; D008565:Membrane Proteins / Q000302:isolation & purification*; D008566:Membranes / Q000648:ultrastructure; D012995:Solubility
D008565:Membrane Proteins