Waters Enzymate BEH Pepsin Columns - Manual

[ CARE AND USE MANUAL ]
CONT ENTS
I. INT RODUCTION
II. HOW WAT ERS HDX T ECHNOLOGY WORKS
III. GET TING START ED
a. Column Connection
b. Column Installation and Equilibration
c. Digesting/Trapping Flow Rate
d. Quenching
e. Protein Injection
f. Trapping Time
g. pH and Solvent Compatibility
h. Temperature
i. Column Washing/Cleaning
j. Column Storage
IV. CHECKING FOR ENZYMATIC ACTIVIT Y
V. REFERENC ES
Enzymate BEH Pepsin Column
I. INT RODUCTION
Thank you for choosing the Waters Enzymate™ BEH Pepsin Column.
Pepsin is immobilized onto rugged 5 μm BEH (Ethylene Bridged
Hybrid) particles which are packed into low dispersion 2.1 x 30 mm
stainless steel column hardware containing 0.5 μm porosity inlet
and outlet frits. This is the same column hardware used in our
eXtended Performance [XP] Column line. This online, flow-through
digestion column reproducibly digests intact proteins into peptides
after incubating with H
2
O/D
2
O during HDX experiments. Benefits
of online protein digestion include reduced sample preparation
time, reproducible digests, minimized back-exchange, and limited
enzyme in solution.
The Enzymate BEH Pepsin Column is housed in a temperature-
controlled compartment to ensure optimal performance and
reproducibility. Once the peptic peptides elute from the pepsin
column, they are focused on a trap column. The peptides are then
separated at 0 ˚C on an ACQUITY UPLC
® BEH C
18
Column and
detected by high resolution mass spectrometry.
The Enzymate BEH Pepsin Column was designed and tested
specifically for use on the ACQUITY UPLC M-Class System with
HDX Technology and the nanoACQUITY UPLC
® System with HDX
Technology. Waters cannot support the use of the Enzymate BEH
Pepsin Column on any other LC system.