Catching up on the latest breakthrough journal articles? Taking stock of reagent requirements for next week’s experiments? Sneaking in a much-needed coffee break?
Frankly, who has the time? With so many samples to prepare, day after day, week after week, month after month, there’s time for little else. And those repetitive tasks of pipetting over and over again, just keep, well, repeating.
It’s time to automate your sample prep! In this technical note, see how researchers reduce day-to-day variability resulting from manual, time-consuming, and multi-step protocols such as the protein denaturation, reduction, alkylation and digestion steps which precede LC/MS analysis. Here, the researchers use a Biomek NXP Span-8 Workstation and protein preparation kits with ready-to-use reagents to automate the entire workflow. Then, they tested the automated workflow for their quantitative proteomics studies.
First, the reproducibility of the digestion protocol alone was tested. As shown in the technical note, very high reproducibility was obtained, with about 90% of peptides monitored by LC/MS having digestion reproducibility below 10% CVs. Next, the digestion protocol was transferred to the Biomek NXP Span-8 Workstation. Many of the proteotypic peptides analyzed showed very high reproducibility with %CVs ~ 2-3%. As expected, for both the manual and automated protocol some peptides showed higher variability than others from the same protein, reflecting inherent digestion heterogeneity. When developing assays for peptide/protein quantitation, it’s important to assess the variability in digestion reproducibility in order to choose the highest performing peptides. Automation of this step can greatly simplify the assessment.
Finally, the researchers tested the reproducibility across different labs and workstations, and across multiple days. The results show very similar cumulative reproducibility curves with more than 80% of peptides monitored having CVs below 10%, demonstrating that the automation method and protocol were very robust and transferable.
To learn more, download the full technical note and see how the Biomek Workstation with optimized protein preparation kits can help you save time while providing reproducible sample preparation today.
Now, what will you do with all that free time?