Waters - Enzymate Online Digestion Columns
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Online Protein Digestion
Waters Enzymate™ BEH Pepsin Column (2.1 x 30 mm, 5 Μm) digests intact proteins into peptides. The peptic peptides are then retained on a trapping column. Peptides elute from the trapping column and are refocused onto a sub-2-Μm ACQUITY UPLC® Column for elution into a high resolution mass spectrometry.
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Features of Enzymate Online Digestion Columns
[u]Column Features[/u]
Waters Enzymate BEH Pepsin Column:
Provides fast, reproducible, and efficient online protein digestion, typically within 30 seconds
Shortens overall protein sample preparation time
Offer the ability to optimize protein digestion efficiency by changing temperature and/or flow rate
[u]Low and High Pressure Digestion Efficiency Comparisons[/u]
Shown on the right is the digestion efficiency comparison between low and high pressure digestions. The Waters Enzymate BEH Pepsin Column was used for digestion pressure at 1,000 psi (low) and 13,500 psi (high). Higher pressure digestion enhanced digestion efficiency.
[u]ACQUITY UPLC M-Class HDX System[/u]
Hydrogen deuterium exchange mass spectrometry (HDX MS) is used to study a protein’s structural dynamics and conformational changes, a component of understanding a protein’s higher order structure. Information about protein conformation from an HDX MS study can be used to compare a control with an analyte by measuring the relative amount of deuteration uptake. HDX MS is capable of monitoring domain interaction, localized protein breathing, and folding or unfolding in the solution phase. The ACQUITY UPLC M-Class System can quantify small changes in protein conformation by extending its pressure range to enable a higher efficiency separation. An integral part of the ACQUITY UPLC M-Class HDX System is the Waters Enzymate BEH Pepsin Column which performs online protein digestion.
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