Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Kwapiszewska, G; Meyer, M; Bogumil, R; Bohle, RM; Seeger, W; Weissmann, N; Fink, L.
Identification of proteins in laser-microdissected small cell numbers by SELDI-TOF and Tandem MS.
BMC Biotechnol. 2004; 4(8):30-30
Doi: 10.1186/1472-6750-4-30
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- Führende Autor*innen der Med Uni Graz
- Kwapiszewska-Marsh Grazyna
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- Abstract:
- BACKGROUND: Laser microdissection allows precise isolation of specific cell types and compartments from complex tissues. To analyse proteins from small cell numbers, we combine laser-microdissection and manipulation (LMM) with mass spectrometry techniques. RESULTS: Hemalaun stained mouse lung sections were used to isolate 500-2,000 cells, enough material for complex protein profiles by SELDI-TOF MS (surface enhanced laser desorption and ionization/time of flight mass spectrometry), employing different chromatographic ProteinChip Arrays. Initially, to establish the principle, we identified specific protein peaks from 20,000 laser-microdissected cells, combining column chromatography, SDS-PAGE, tryptic digestion, SELDI technology and Tandem MS/MS using a ProteinChip Tandem MS Interface. Secondly, our aim was to reduce the labour requirements of microdissecting several thousand cells. Therefore, we first defined target proteins in a few microdissected cells, then recovered in whole tissue section homogenates from the same lung and applied to these analytical techniques. Both approaches resulted in a successful identification of the selected peaks. CONCLUSION: Laser-microdissection may thus be combined with SELDI-TOF MS for generation of protein marker profiles in a cell-type- or compartment-specific manner in complex tissues, linked with mass fingerprinting and peptide sequencing by Tandem MS/MS for definite characterization.
- Find related publications in this database (using NLM MeSH Indexing)
- Animals -
- Biological Markers - metabolism
- Chromatography, Affinity - methods
- Electrophoresis, Polyacrylamide Gel - methods
- Frozen Sections - methods
- Lung - chemistry Lung - cytology
- Male -
- Mass Spectrometry - methods
- Mice -
- Mice, Inbred BALB C -
- Microdissection - methods
- Peptide Fragments - chemistry Peptide Fragments - metabolism
- Peptide Mapping - methods
- Protein Array Analysis - methods
- Proteins - chemistry
- Pulmonary Alveoli - chemistry Pulmonary Alveoli - cytology
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods