Source:http://linkedlifedata.com/resource/pubmed/id/16307718
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
2006-1-24
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pubmed:abstractText |
A small amount of bioptic tissue ( approximately 5-10mg of fresh tissue) usually does not contain enough material to extract protein and RNA separately, to obtain preparative two-dimensional polyacrylamide gel electrophoresis (2-DE), and to identify a large number of separated proteins by MS. We tested a method, on small renal cancer specimens, for the coextraction of protein and RNA coupled with 2-DE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) or quadrupole time-of-flight (Q-TOF) analysis. We coextracted 0.28+/-0.05mg of proteins and 2.5+/-0.33microg of RNA for each 10mg of renal carcinoma tissue. Small and large 2-DE gels were compared: they showed a similar number of spots, and it was possible to match each other; using small format gels, one-fifth of the protein amount was required to identify, by Q-TOF analysis, the same number of proteins identifiable in large-format gel using MALDI-TOF analysis. Quality of RNA coextracted with the proteins was tested by real-time PCR on a set of housekeeping genes. They were quantified with high amplification efficiency and specificity. In conclusion, using 5 to 10mg of fresh tissue, it was possible to perform comprehensive parallel proteomic and genomic analysis by high-resolution, small-format 2-DE gels, allowing approximately 300 proteins identification and 1000 genes expression analysis.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
0003-2697
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
349
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
62-71
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pubmed:meshHeading |
pubmed-meshheading:16307718-Carcinoma, Renal Cell,
pubmed-meshheading:16307718-Electrophoresis, Gel, Two-Dimensional,
pubmed-meshheading:16307718-Gene Expression Profiling,
pubmed-meshheading:16307718-Humans,
pubmed-meshheading:16307718-Kidney Neoplasms,
pubmed-meshheading:16307718-Neoplasm Proteins,
pubmed-meshheading:16307718-Polymerase Chain Reaction,
pubmed-meshheading:16307718-Proteomics,
pubmed-meshheading:16307718-RNA, Neoplasm,
pubmed-meshheading:16307718-Spectrometry, Mass, Matrix-Assisted Laser...
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pubmed:year |
2006
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pubmed:articleTitle |
Protein/RNA coextraction and small two-dimensional polyacrylamide gel electrophoresis for proteomic/gene expression analysis of renal cancer biopsies.
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pubmed:affiliation |
Dipartimento di Genetica, Biologia, Biochimica-Università degli Studi di Torino, 10126 Torino, Italy. giovanna.barbero@unito.it
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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