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A Simple Protein Extraction Method for Proteomic Analysis of Diverse Biological Specimens

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Title A Simple Protein Extraction Method for Proteomic Analysis of Diverse Biological Specimens
 
Creator REDDY, PJ
RAO, AA
MALHOTRA, D
SHARMA, S
KUMAR, R
JAIN, R
GOLLAPALLI, K
PENDHARKAR, N
RAPOLE, S
SRIVASTAVA, S
 
Subject 2-DE
2D-DIGE
breast cancer
B. subtilis
glioblastoma cell line
mass spectrometry
protein extraction
S. coelicolor
TRIzol
yeast
2-DIMENSIONAL GEL-ELECTROPHORESIS
STREPTOMYCES-COELICOLOR
SACCHAROMYCES-CEREVISIAE
BACILLUS-SUBTILIS
GLIOBLASTOMA-MULTIFORME
SAMPLE PREPARATION
BREAST-CANCER
HUMAN GLIOMAS
IDENTIFICATION
RNA
 
Description The success of a proteomic experiment largely depends on the quality and quantity of the protein extract. Currently, various protocols are available for extraction of proteins from different types of samples; however, further optimization is required for every new sample type. Hence, a common protein extraction protocol is desirable. In the present study, soluble proteins were extracted from six diverse samples using TRIzol without any additional clean-up step and subjected to 2-DE and 2D-DIGE analysis for global protein expression profiling. Image analysis using IMP7 and DeCyder showed good coverage, reproducibility and quality of the gel. MS analysis of 24 spots from all the six samples showed good score and coverage for the identified proteins. Additionally, this method facilitated the concurrent isolation of RNA from the same cell lysates with high integrity and quality, suitable for transcriptomic analysis. Thus, we demonstrate the use of a common protein extraction protocol involving TRIzol reagent for 2-DE, 2D-DIGE and MS analysis using six diverse samples and show its suitability for concomitant transcriptomic studies.
 
Publisher BENTHAM SCIENCE PUBL LTD
 
Date 2014-10-15T11:03:16Z
2014-10-15T11:03:16Z
2013
 
Type Article
 
Identifier CURRENT PROTEOMICS, 10(4)298-311
1570-1646
1875-6247
http://dx.doi.org/10.2174/15701646113106660004
http://dspace.library.iitb.ac.in/jspui/handle/100/14814
 
Language en