Tuesday, December 26, 2017
Impact of detergents on membrane complex isolation!
Ever needed a good protocol to analyze a protein complex with some membrane components? Check out this awesome systematic analysis!
This team focuses on something called Cad11 (it appears to have some really interesting significance in cancer on top of having a membrane linked protein complex structure appropriate to exactly this type of assay optimization.
There are some really striking differences in the study including when the use of one detergent leads to the complete loss of the ability to pull down their complex(!!) when others can as well as the observation of novel proteins only observed when certain detergents are utilized. Sample prep optimizatin, FTW!
iTRAQ 4-plex reagent is used for a quantitative readout of complex isolation efficiency and is analyzed on an Orbitrap Fusion Lumos system using MS2 based quantification.
Considering how important the different detergents appear to be here, it might be fair to wonder if these results would reproduce for different protein complexes.... If this isn't your protein of interest, it at least shows you how valuable picking the right detergents can be and gives you a good starting point!