Document Detail


Protein Analysis Through Western Blot of Cells Excised Individually from Human Brain and Muscle Tissue.
MedLine Citation:
PMID:  22402104     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Comparing protein levels from single cells in tissue has not been achieved through Western blot. Laser Capture Microdissection allows for the ability to excise single cells from sectioned tissue and compile an aggregate of cells in lysis buffer. In this study we analyzed proteins from cells excised individually from brain and muscle tissue through Western blot. After we excised individual neurons from the substantia nigra of the brain, the accumulated surface area of the individual cells added to 120,000, 24,000, 360,000, 480,000, 600,000 μm(2). We used an optimized Westerm blot protocol to probe for tyrosine hydroxylase in this cell pool. We also took 360,000 μm(2) of astrocytes (1700 cells) and analyzed the specificity of the method. In muscle we were able to analyze the proteins of the five complexes of the electron transport chain through Western Blot from 200 human cells. With this method, we demonstrate the ability to compare cell specific protein levels in the brain and muscle and describe for the first time how to visualize proteins through Western Blot from cells captured individually.
Authors:
A O Koob; L Bruns; C Prassler; E Masliah; T Klopstock; A Bender
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-3-5
Journal Detail:
Title:  Analytical biochemistry     Volume:  -     ISSN:  1096-0309     ISO Abbreviation:  -     Publication Date:  2012 Mar 
Date Detail:
Created Date:  2012-3-9     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012 Elsevier Inc. All rights reserved.
Affiliation:
Department of Neurology, Klinikum Grosshadern, University of Munich, Marchinionistr. 23, 81377 Munich, Germany.
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