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Analysis of protein and Peptide profiling of body fluids by surface-enhanced laser desorption/ionization mass spectrometry (CAT#: STEM-ST-0341-LJX)

Introduction

Recently, Ciphergen Biosystems (Fremont, CA, USA) has developed a technique called surface-enhanced laser desorption/ionization (SELDI). This technology is based on ProteinChips(R) with chemically or biochemically modified surfaces for the selective retention and enrichment of protein subsets from a complex protein mixture. The proteins or peptides of interest can then be identified by using mass spectrometry (SELDI-MS). This highly sensitive and high-throughput technique can detect minute differences in proteins and peptide profiles between biological samples. The versatility of this technology enables a wide variety of applications in basic research, clinical, proteomic, and drug discovery such as identification of novel biomarkers associated with certain diseases or treatments. Many disease biomarkers or biomarker patterns have been identified using SELDI-MS in various laboratories.




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:
Body fluids

Notes

When operating, strictly follow the experimental steps.