Waters Sirocco Protein Precipitation Plate - Manual (Page 2)

[ Care and Use ManUal ]
2 Sirocco Protein Precipitation Plate
Step 4 – Seal
Apply vented cap mat, using a cap roller to ensure secure, uniform sealing.
This will prevent splashing and cross-talk during mixing.
Cap Mat
Step 5 – Mix
Shake (mix) the loaded Sirocco plate/collection plate assembly on a vortexer
(e.g., Multi-Tube Vortexer from VWR) at medium setting (e.g., setting 5 on a
1-10 scale) for 0.5 to 1.0 minute.
Note: There is no need to remove the vented cap mats or valve tips from the
Sirocco Plate before filtration. This eliminates the risk of cross-talk between
wells. The valves and vented cap mat are designed to open under vacuum
to allow controlled flow during filtration.
Note: Excessive mixing at too high a setting (e.g., setting 8 on a 1-10
scale) could cause cross-talk. Under mixing (e.g., setting 2 on a 1-10 scale)
will cause clogging or a cloudy filtrate.
Step 6 – Filter
After mixing, place the Sirocco plate on a Vacuum Manifold (e.g. Waters
vacuum manifold P/N 186001831), and apply 8-5” Hg (271-508 mbar)
of vacuum for up to three minutes. A higher vacuum setting can be used if
flow does not occur at 10” Hg.
Note: The Sirocco proprietary valve tip design prevents liquid flow through
the device until a minimum 5” Hg (169 mbar) of vacuum is applied.
Note: The Sirocco Plate can be filtered by using centrifiguation (@ 2,000 x g
for 1 min.) in place of vacuum device.
Step 7– Analysis
Discard the Sirocco Plate and remove the collection plate containing clear
filtrate from the vacuum device, seal and transfer to the LC/MS autosam-
pler or store for later analysis.
Note: There is no need to remove cap mats or valve tips from Sirocco Plate.
The Sirocco Plate is discarded after filtration.
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