| Decreased sensitivity associated with an altered formulation of a commercially available kit for detection of protein carbonyls. | |
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MedLine Citation:
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PMID: 20230891 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Carbonylation is a commonly studied form of oxidative modification to proteins which can be conveniently detected using commercially available kits. The most common of these kits is the Oxyblot Protein Oxidation Detection Kit (Chemicon/Millipore). Over the past year we have observed severely diminished sensitivity of these kits which was shown to be a result of a change in the formulation of one of the components supplied in the kit. This component, the 10X 2,4-dinitrophenylhydrazine derivatization solution, which had previously been dissolved in 100% trifluoroacetic acid (TFA), was now dissolved in 2N hydrochloric acid, which according to our results is not acid enough. Further, we observed that upon storage even DNPH dissolved in TFA is subject to degradation. Based on these studies, we make recomendations that should improve the sensitivity and reproducibilty of this assay. |
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Authors:
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Ping Wang; Saul R Powell |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2010-03-12 |
Journal Detail:
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Title: Free radical biology & medicine Volume: 49 ISSN: 1873-4596 ISO Abbreviation: Free Radic. Biol. Med. Publication Date: 2010 Jul |
Date Detail:
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Created Date: 2010-08-04 Completed Date: 2011-04-19 Revised Date: 2011-08-01 |
Medline Journal Info:
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Nlm Unique ID: 8709159 Medline TA: Free Radic Biol Med Country: United States |
Other Details:
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Languages: eng Pagination: 119-21 Citation Subset: IM |
Copyright Information:
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Copyright 2010 Elsevier Inc. All rights reserved. |
Affiliation:
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Cardiac Metabolism Laboratory, Center for Heart and Lung Research, The Feinstein Institute for Medical Research, Manhasset NY, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Biochemistry / methods Complex Mixtures / chemistry* Fluorenes / chemistry* Humans Hydrazines / chemistry* Hydrochloric Acid / chemistry* Hydrogen-Ion Concentration Protein Carbonylation Reagent Kits, Diagnostic* Reproducibility of Results Sensitivity and Specificity |
| Grant Support | |
ID/Acronym/Agency:
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HL68936/HL/NHLBI NIH HHS; R01 HL068936-06/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Complex Mixtures; 0/Fluorenes; 0/Hydrazines; 0/Reagent Kits, Diagnostic; 0/dinitrophenylhydrazine; 0/trifluorenylamine; 7647-01-0/Hydrochloric Acid |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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