Waters MassPREP Phosphopeptide Enrichment Kit User Manual

Care and use manual, Massprep, Phosphopeptide enrichment kit

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[ Care and Use ManUal ]

I. Int roductIon

The MassPREP

Phosphopeptide Enrichment Kit is a tool for selective

enrichment of phosphopeptides from complex proteomic samples. This
kit contains a micro-scale solid-phase extraction (SPE) plate packed with
a unique affinity sorbent. This affinity SPE does not require pre-loading
the sorbent with metal chelation. Also, superior selectivity is achieved
compared to other existing affinity chromatographic methods.

The selectivity of this sorbent towards phosphopeptides is further
improved by adding a unique chemical to the sample before loading
onto the SPE. This chemical (under the trade name, Enhancer) improves
the overall selectivity by displacing the non-phosphorylated peptides
during the sample loading step.

The MassPREP

Phosphopeptide Enrichment Kit includes the 96-well

micro elution (µElution) plate, enhancer, and a test sample for method
validation. The test sample is a mixture of four synthetic phosphopeptides
mixed with yeast enolase tryptic peptides in 1:1 molar ratio. A general
guideline for kit operation and specific protocols for phosphopeptide
extraction from complex protein digests are provided. The 96-well
micro elution (µElution) platform conveniently uses a vacuum manifold,
for reproducibility and high throughput of enrichment experiments.

contEntS

I. INTRODUCTION

II. MATERIALS INCLUDED IN THE KIT

III. PHOSPHOPEPTIDE AFFINITY ENRICHMENT METHOD

IV. PHOSPHOPEPTIDE ENRICHMENT KIT USAGE GUIDELINES

Part A. How to Use the MassPREP

Phosphopeptide Enrichment

μElution Plate

Part B. How to Use the MassPREP

Enolase/Phosphopeptide

Standard as a Test Sample

Part C. How to Apply Enhancer for Highly Complex Samples

Part D. Sample Preparation for Mass Spectrometry Analysis

V. ORDERING INFORMATION

VI. STORAGE AND DISPOSAL OF USED PLATES

MassPrEP

PhoSPhoPEPtIdE EnrIchMEnt KIt

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