Unleash the power of DIA
Available on demand
In the ever lasting effort to boost the number of protein identified in MS-based proteomic experiments, it has become clear that Data Independent Analysis (DIA), combined with short gradients, has become a winning combination.
The steep development over the last years, have positioned DIA as an important acquisition strategy to generate record high identifications with short gradients on the Evosep One.
Join this webinar to explore how our customers have used Evosep One to generate record high IDs per minute for large-scale sample cohorts. You can also read more about the research published using DIA methodologies from our users here.
Rapid and in-depth coverage of the (phospho-)proteome with deep libraries and optimal window design for dia-PASEF
Patricia Skowronek, PhD student, DEPARTMENT OF PROTEOMICS AND SIGNAL TRANSDUCTION, MAX PLANCK INSTITUTE OF BIOCHEMISTRY
Data-independent acquisition combined with parallel accumulation ‒ serial fragmentation (dia-PASEF) uses the correlation of the mass to charge ratio to the ion mobility separation in a Bruker timsTOF mass spectrometer.
In this webinar, I will elaborate on how to generate dia-PASEF methods with variable isolation widths and optimal isolation window placement. Additionally, I will demonstrate that these optimal methods applied to short Evosep gradients with project-specific in-depth libraries lead to a deep proteome and even deeper phosphoproteome. In only 21 min (60 per day/method), we quantify the HeLa cell proteome to a depth of more than 7,000 and the phosphoproteome to more than 10,000 distinct class I phosphosites. I will also show advanced visualization of these results in the AlphaMap and AlphaViz environment.
Combining Evosep One with state of the art SWATH acquisition
Talk by Yves Le Blanc, Associate Chief Research Scientist, SCIEX, and
Talk by Ihor Batruch, Technical Product Manager at Sciex
Accurate and reproducible compound identification from complex mixtures is ultimately the goal of any LC-MSMS analysis. To achieve this on a large number of analytes, Data Independent Analysis (DIA) methods have emerged as MSMS technique that can reliably detect the response of analytes over their entire chromatographic peak. Combination of these techniques with high performance LC separation provides higher confidence in compound identification.
Here we present the combination of rapid LC analysis provided by the Evosep system, along with SWATH acquisition approaches. The first approach focussed on Scanning-SWATH, over small mass range, in combination with the Evosep workflow of 200 and 100spd. Using 1 amu window and fast sample turn around, maximum coverage can be achieved with high confidence through replicate analysis.
The second approach focused on protein identification and quantification achievable with a ZenoTOF 7600 system in SWATH acquisition mode coupled to Evosep operated at workflows of 200, 100, 60 and 30 SPD across different HeLa digest sample loads. Performance improvements of a novel Zeno SWATH acquisition strategy relative to SWATH acquisition on a prototype ZenoTOF 7600 system will also be evaluated. A preliminary look at how these combined technologies can improve sample throughput as well compound coverage with be presented.