Waters MassPREP Phosphopeptide Enrichment Kit - Manual (Page 4)

[ Care and Use ManUal ]

Part C - How to Apply MassPREP ™ Enhancer for Highly
Complex Samples
MassPREP ™ Enhancer can be used to improve the overall selectivity
of the phosphopeptides from a highly complex biological sample (e.g.,
whole cell lysate digested with proteolytic enzymes).
1. Weigh 100 mg Enhancer from the vial and transfer to a separate
vial. Keep the rest of Enhancer in the capped vial for later use, and
store the vial at room temperature. Expiration time of the Enhancer
powder is ~ 3 years; storage is at room temperature.
2. Prepare 100 mg/mL Enhancer solution by adding 1 mL of the loading
solution to the weighed-out enhancer, vortex gently. (Note: The
expiration time for this reconstituted solution is ~ 1 week; storage
is at 4 ˚C)
3. Prepare the peptide sample by solubilizing it with 200 µL of the
Enhancer solution.
4. Precede with Part I to isolate the phosphopeptides.
Part D - Sample Preparation for Mass Spectrometry Analysis
The phosphopeptides enriched using this kit, will be subjected to Mass
Spectrometry analysis. Here are some advices for post-kit sample
preparation.
1. For MALDI-MS analysis, use a reversed-phase cartridge (e.g, ZipTip ™ )
to remove Diammonium Phosphate or other potential contaminants
from the phosphopeptides.
2. For capillary or nano-scale LC/MS analysis, a reversed-phase trap
column is needed to remove excess EDTA, Diammonium Phosphate
and other contaminants from the sample.
V. ordErIng InforMatIon
Table 2. MassPREP ™ Phosphopeptide Enrichment Kit Ordering Information
Description Part Number Units/ki Amt/Unit
MassPREP ™ Phosphopeptide Enrichment Kit 186003864 – –
MassPREP ™ Phosphopeptide Enrichment
µElution Plate
186003820 Qty 1 1 plate
MassPREP ™ Enolase/Phosphopeptide
Standard
186003286 Qty 1 1 nmole
MassPREP ™ Enhancer ™ 186003821 Qty 10 500mg
Table 3. Vacuum Manifold Accessories Ordering Information
SPE Vacuum Pump (115, 60 Hz) 725000417
SPE Vacuum Pump (240, 50 Hz) 725000418
µElution Plate Manifold 186001831
Manifold Waste Tray, disposable, 25/pkg WAT058942
350 µL Sample Collection Plate, polypropylene
96 wells, 50 plates/pkg WAT058943
VI. StoragE and dISPoSal of uSEd PlatES
Cartridges stored in their original sealed pouch remain stable for long
periods. To store unused cartridges in opened pouches, squeeze the
air out of the pouch, fold over the open end of the pouch twice, seal
with tape and store in a dessicator. Contaminants that may interfere
with the effectiveness of the affinity sorbent are high quantities of
detergents and organic salts, such as urea. Dispose off used cartridges
and plates safely in accordance with applicable government or local
regulations.