Linked Life Data

20373496

Source:http://linkedlifedata.com/resource/pubmed/id/20373496

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2010-4-7
pubmed:abstractText
The Click-iT EdU cell proliferation assay (Invitrogen) for detection of replicating cells is based on incorporation of EdU into newly synthesized DNA and its recognition by azide dyes via a copper mediated "click" reaction. In the protocol provided by Invitrogen, cells are fixed with paraformaldehyde and stained with 7-aminoactinomycin D (7-AAD) for DNA content analysis. Both of these procedures result in DNA histograms with a broad coefficient of variation. We have modified this protocol and show that after EdU incorporation, nuclei isolated by hypotonic lysis of cells can be directly labeled using the Click-iT Alexa Fluor 488 Assay kit and stained with propidium iodide. This modified procedure using isolated nuclei and propidium iodide staining results in DNA histograms with better resolution (lower coefficient of variation of the G(1) peak) and shorter processing time by eliminating the fixation and permeabilization steps.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
1934-9300
pubmed:author
pubmed:copyrightInfo
(c) 2010 by John Wiley & Sons, Inc.
pubmed:issnType
Electronic
pubmed:volume
Chapter 7
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
Unit7.36
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
Click-iT proliferation assay with improved DNA histograms.
pubmed:affiliation
University of Miami Miller School of Medicine, Miami, Florida, USA.
pubmed:publicationType
Journal Article