PMID- 17141588
OWN - NLM
STAT- MEDLINE
DCOM- 20070611
LR  - 20070410
IS  - 1570-0232 (Print)
IS  - 1570-0232 (Linking)
VI  - 849
IP  - 1-2
DP  - 2007 Apr 15
TI  - Disease proteomics of high-molecular-mass proteins by two-dimensional gel 
      electrophoresis with agarose gels in the first dimension (Agarose 2-DE).
PG  - 211-22
AB  - Agarose gel is the preferred electrophoretic medium currently used for separating 
      high molecular mass (HMM) proteins (MW>100 kDa). Agarose gels are widely used for 
      both SDS-agarose gel electrophoresis and agarose isoelectric focusing (IEF). A 
      two-dimensional gel electrophoresis method employing agarose gels in the first 
      dimension (agarose 2-DE) that is sufficiently good at separating up to 1.5mg of 
      HMM proteins with molecular masses as large as 500 kDa has been used to separate 
      proteins from various diseased tissues and cells. Although resolution of the 
      agarose 2-DE pattern always depends on the tissue being analyzed, sample 
      preparation procedures including (i) protein extraction with an SDS sample 
      buffer; (ii) ultracentrifugation of a tissue homogenate; and (iii) 1% SDS in both 
      stacking and separation gels of the second-dimension SDS-PAGE gel, are generally 
      effective for HMM protein detection. In a comprehensive prostate cancer proteome 
      study using agarose 2-DE, the HMM region of the gel was rich in proteins of 
      particular gene/protein expression groups (39.1% of the HMM proteins but only 
      28.4% of the LMM ones were classified as transcription/translation-related 
      proteins). Examples include transcription factors, DNA or RNA binding proteins, 
      and ribosomal proteins. To understand oxidative stress-induced cellular damage at 
      the protein level, a novel proteomic method, in which protein carbonyls were 
      derivatized with biotin hydrazide followed by agarose 2-DE, was useful for 
      detecting HMM protein carbonyls in tissues of both a diabetes model Ostuka 
      Long-Evans Tokushima Fatty (OLETF) rat and a control Long-Evans Tokushima Otsuka 
      (LETO) rat. In this paper, we review the use of agarose gels for separation of 
      HMM proteins and disease proteomics of HMM proteins in general, with particular 
      attention paid to our proteome analyzes based on the use of agarose 2-DE for 
      protein separation followed by the use of mass spectrometry for protein 
      identification.
FAU - Oh-Ishi, Masamichi
AU  - Oh-Ishi M
AD  - Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University 
      School of Science, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan. 
      oishi@walrus.sci.kitasato-u.ac.jp
FAU - Maeda, Tadakazu
AU  - Maeda T
LA  - eng
PT  - Journal Article
PT  - Review
DEP - 20061201
PL  - Netherlands
TA  - J Chromatogr B Analyt Technol Biomed Life Sci
JT  - Journal of chromatography. B, Analytical technologies in the biomedical and life 
      sciences
JID - 101139554
RN  - 0 (Proteins)
SB  - IM
MH  - Animals
MH  - *Disease
MH  - Electrophoresis, Agar Gel/*methods
MH  - Electrophoresis, Gel, Two-Dimensional/*methods
MH  - Humans
MH  - Molecular Weight
MH  - Proteins/*analysis/chemistry
MH  - Proteomics/*methods
RF  - 61
EDAT- 2006/12/05 09:00
MHDA- 2007/06/15 09:00
CRDT- 2006/12/05 09:00
PHST- 2006/06/16 00:00 [received]
PHST- 2006/09/30 00:00 [revised]
PHST- 2006/10/27 00:00 [accepted]
PHST- 2006/12/05 09:00 [pubmed]
PHST- 2007/06/15 09:00 [medline]
PHST- 2006/12/05 09:00 [entrez]
AID - S1570-0232(06)00888-9 [pii]
AID - 10.1016/j.jchromb.2006.10.064 [doi]
PST - ppublish
SO  - J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Apr 15;849(1-2):211-22. doi: 
      10.1016/j.jchromb.2006.10.064. Epub 2006 Dec 1.