Comparative proteomic analysis using samples obtained with laser microdissection and saturation dye labelling

Comparative proteomic methods are rapidly being applied to many different biological systems including complex tissues. One pitfall of these methods is that in some cases, such as oncology and neuroscience, tissue complexity requires isolation of specific cell types and sample is limited. Laser micr...

Full description

Saved in:
Bibliographic Details
Published in:Proteomics (Weinheim) 2005-10, Vol.5 (15), p.3851-3858
Main Authors: Wilson, Kate E., Marouga, Rita, Prime, John E., Pashby, D. Paul, Orange, Paul R., Crosier, Steven, Keith, Alexander B., Lathe, Richard, Mullins, John, Estibeiro, Peter, Bergling, Helene, Hawkins, Edward, Morris, Christopher M.
Format: Article
Language:English
Subjects:
2-DE
Alzheimer Disease - genetics
Alzheimer Disease - pathology
Alzheimer's disease
Amyloid beta-Protein Precursor - genetics
Amyloid beta-Protein Precursor - metabolism
Analytical, structural and metabolic biochemistry
Animals
Animals, Genetically Modified
Biological and medical sciences
Coloring Agents
Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases
Disease Models, Animal
Electrophoresis, Gel, Two-Dimensional - methods
Fundamental and applied biological sciences. Psychology
Hippocampus - chemistry
Hippocampus - metabolism
Image Processing, Computer-Assisted
Laser microdissection
Lasers
Male
Medical sciences
Microdissection - methods
Miscellaneous
Neurology
Proteins
Proteins - chemistry
Proteins - genetics
Proteins - isolation & purification
Proteomics - methods
Pyramidal Cells - chemistry
Rats
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
Transgenics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Comparative proteomic methods are rapidly being applied to many different biological systems including complex tissues. One pitfall of these methods is that in some cases, such as oncology and neuroscience, tissue complexity requires isolation of specific cell types and sample is limited. Laser microdissection (LMD) is commonly used for obtaining such samples for proteomic studies. We have combined LMD with sensitive thiol‐reactive saturation dye labelling of protein samples and 2‐D DIGE to identify protein changes in a test system, the isolated CA1 pyramidal neurone layer of a transgenic (Tg) rat carrying a human amyloid precursor protein transgene. Saturation dye labelling proved to be extremely sensitive with a spot map of over 5,000 proteins being readily produced from 5 μg total protein, with over 100 proteins being significantly altered at p 
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200401255