Purifications of multiprotein complexes are challenging and typically more difficult than that of individual proteins. Many complexes are quite fragile and in general, a larger number of purification steps may be needed to achieve the desired purity.
Given the complexity of maintaining a stable, intact and functional complex until the end of the purification process, it’s key to have a precise technology that monitors the quality and functionality of the collected fractions at any step during the purification workflow. This allows you to make better decisions on whether to move forward, without wasting time and effort on questionable samples that lead only to inconclusive and non-reproducible results.
Identifying the fractions containing your functional complex in minutes
Standard methods to monitor complex formation are usually time and sample-consuming, including SDS-PAGE and western blots. Moreover, its functionality must be assessed in separate assays, complicating even further the overall process.
In this application note, learn how the Tycho NT.6 system is used throughout the purification of the chromatin remodeler complex HSAX to monitor its formation and functionality in the same experiment. Using only a small amount of sample, Tycho is able to unambiguously identify the complex-containing fractions using a thermal unfolding profile and also to test its functionality by binding interactions while shrinking analyses time from hours to minutes.