Free sex speed dates - Tracking and elucidating alphavirus host protein interactions
Recently the use of multiple tags, such as the two-step tandem affinity purification tag strategy, has become popular (8).Although all these tags have proved highly effective for the isolation of protein complexes, they cannot be readily visualized in living cells.
Two-liter cultures were grown at 37 °C in LB medium (Fisher) with ampicillin until EDTA, 0 protease inhibitors (16) to make up to a total of 200 ml).
Cells were lysed using an M110S microfluidizer (Microfluidics International Corp., Newton, MA).
Localization of such complexes provides key insights into their functions.
There are a variety of tools for localizing individual proteins in space and time in the cellular milieu (1).
We applied this method to the analysis of a variety of endogenous protein complexes from different eukaryotic cells.