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Revealing the Significant Artifacts in Serum Obtained from Clot Activator–Containing Collection Devices by Surface-Enhanced Laser Desorption Ionization/Time-of-Flight Mass Spectrometry (CAT#: STEM-ST-0334-LJX)

Introduction

Variations in specimen collection and processing may confound analyses of protein profiles obtained by mass spectrometry. Recently, Banks et al. demonstrated that the choice of device for specimen collection affected the observed plasma proteomic profile. Extending these observations, we observed alteration of the mass spectrometric protein profile for specimens collected in clot activator– and gel-containing tubes, compared with specimens collected in the plain tubes of Banks et al.




Principle

The surface enhanced laser desorption ionization technique belongs to laser desorption mass spectrometry (LDMS). It is different from ordinary LDMS in that the laser is not directly hit on the sample to desorption, but the sample is suspended in the matrix, the laser is hit on the matrix, the matrix absorbs and transmits the laser energy, so that the sample in the matrix desorption out. After desorption and ionization, the samples were examined in a time-flight mass spectrometer.

Applications

For protein analysis and measurement of molecular weight of complete proteins
For the diagnosis of a variety of diseases, especially cancer

Procedure

1. The surface of the protein chip is treated in a certain chemical or biochemical way (surface enhancement), so that it has the ability to bind specifically to a certain type of protein
2. The serum or protein extract is directly added to the surface of the chip, and the chip is washed after incubation. Specific proteins bind to the chip and are thus separated from the protein mixture
3. The chip then uses a "chip reader" (a kind of SELDI-TOF-MS) to obtain a mass spectrum of the protein bound to the chip
4. The SELDI protein chip system can be used to compare changes in the protein profile of any set of control samples or different disease states to identify biomarkers or disease-related targets

Materials

• Sample Type:
Serum

Notes

When operating, strictly follow the experimental steps.
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