Source:http://linkedlifedata.com/resource/pubmed/id/14628683
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
5
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pubmed:dateCreated |
2003-11-21
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pubmed:abstractText |
Using a bisubstituted caspase-3 target sequence: aspartate-glutamate-valine-aspartate, (z-DEVD)2 peptide derivative of the fluorophore, cresyl violet, we have obtained a cell permeant, fluorogenic, caspase substrate capable of detecting the site-specific presence of functionally active, caspase-3 and caspase-7 up-regulation within intact apoptotic cells. Addition of this substrate to induced and noninduced cell culture populations allows for the rapid site-specific detection of caspase up-regulation without the requirement for a wash step. We demonstrate here the use of (z-DEVD)2-cresyl violet substrate for the detection of apoptosis induction in Jurkat, THP-1, and MCF-7 cells using fluorescence microscopy and 96-well fluorescence plate reader analysis. Intracellular up-regulated DEVDase activity, which was clearly visible by fluorescence microscopy and 96-well fluorescence plate reader measurements, showed greater than 6-fold increases in fluorescence output in induced versus noninduced Jurkat cell samples. A simple fluorogenic substrate conversion method is demonstrated here for detecting apoptosis induction within intact living cells.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DEVDase,
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescent Dyes,
http://linkedlifedata.com/resource/pubmed/chemical/Oligopeptides,
http://linkedlifedata.com/resource/pubmed/chemical/Oxazines,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Hydrolases
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0736-6205
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
35
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1080-5
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pubmed:dateRevised |
2010-12-3
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pubmed:meshHeading |
pubmed-meshheading:14628683-Apoptosis,
pubmed-meshheading:14628683-Breast Neoplasms,
pubmed-meshheading:14628683-Cell Count,
pubmed-meshheading:14628683-Cell Line, Tumor,
pubmed-meshheading:14628683-Cell Membrane Permeability,
pubmed-meshheading:14628683-Fluorescent Dyes,
pubmed-meshheading:14628683-Humans,
pubmed-meshheading:14628683-Jurkat Cells,
pubmed-meshheading:14628683-Microscopy, Fluorescence,
pubmed-meshheading:14628683-Oligopeptides,
pubmed-meshheading:14628683-Oxazines,
pubmed-meshheading:14628683-Peptide Hydrolases,
pubmed-meshheading:14628683-Sensitivity and Specificity,
pubmed-meshheading:14628683-Staining and Labeling,
pubmed-meshheading:14628683-Tissue Distribution
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pubmed:year |
2003
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pubmed:articleTitle |
DEVDase detection in intact apoptotic cells using the cell permeant fluorogenic substrate, (z-DEVD)2-cresyl violet.
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pubmed:affiliation |
Immunochemistry Technologies, LLC, Bloomington, MN, USA. brian@immunochemistry.com
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pubmed:publicationType |
Journal Article,
Evaluation Studies
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