Analysis of urine protein by surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry (CAT#: STEM-ST-0353-LJX)

Introduction

In the last few years there has been an increasing interest in exploring the human proteome. In particular, efforts have focused on developing strategies to generate reproducible protein maps of normal cells, tissues, and biologic fluids, from which studies can then compare protein expression between different groups (e.g., healthy individuals vs. those with a specific pathologic state). Various extrinsic factors (instrument settings, matrix composition, urine storage post void, freeze-thaw cycles) and intrinsic factors (blood in urine, urine dilution, first-void vs. midstream urine) were analyzed with respect to their impact on urine protein profiling using surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS).

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Principle Applications Procedure Materials Notes Related Products

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:
Urine

Notes

When operating, strictly follow the experimental steps.

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