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| Institute: |
Lab or Branch |
| The Johns Hopkins University |
Otolaryngology-Head and Neck Surgery, The
Johns Hopkins University |
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| Title: |
| Separation and identification of membrane
proteins from rat cardiac and skeletal |
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| Authors: |
| Z.B. Li, M. Lehar, P.W. Flint |
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| Abstract: |
| The proteomics approaching membrane proteins
is an important initial step in searching for novel target
marker of gene therapy. The membrane protein fractions
were extracted by using Sigma’s membrane extraction
kit. The proteins were separated by using first-dimensional
electrophoresis of immobilized pH 5-8 gradient strips
and following secondary dimensional electrophoresis of
2% SDS-page gel separating gel. The gels were stained
by silver. PDQuest analysis software was applied to locate,
quantities and match spots of individual gels. The distribution
characteristics of the protein expression in both cardiac
and skeletal muscles were analyzed in this study according
to spot molecular, pI and density. Total 96 proteins were
identified in both muscle tissues by Matrix-assisted laser
desorption/ionization-mass spectrometry (MALDI-MS) on
the basis of peptide mass fingerprinting following in
gel digestion with trypsin. Two different methods of extraction
membrane proteins were also comprised in the study. This
result provides a useful reference for comparative studies
of membrane protein of cardiac and skeletal muscles. |
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