A fluorescent probe which allows highly specific thiol labeling at low pH

Publication: Research - peer-reviewJournal article – Annual report year: 2012

View graph of relations

Determination of the thiol-disulfide status in biological systems is challenging as redox pools are easily perturbed during sample preparation. This is particularly pertinent under neutral to mildly alkaline conditions typically required for alkylation of thiols. Here we describe the synthesis and properties of a thiol-specific reagent, fluorescent cyclic activated disulfide (FCAD), which includes the fluorescein moiety as fluorophore and utilizes a variation of thiol-disulfide exchange chemistry. The leaving-group character of FCAD makes it reactive at pH 3, allowing modification at low pH, limiting thiol-disulfide exchange. Different applications are demonstrated including picomolar thiol detection, determination of redox potentials, and in-gel detection of labeled proteins.
Original languageEnglish
JournalAnalytical Biochemistry
Volume421
Issue number1
Pages (from-to)115-120
ISSN0003-2697
DOIs
StatePublished - 2012
Peer-reviewedYes
CitationsWeb of Science® Times Cited: 4

Keywords

  • SDS–PAGE, Fluorescence, Cysteine, Oxidation, In-gel detection, Proteins, Disulfide, Thiol modification, Thiol
Download as:
Download as PDF
Select render style:
APAAuthorCBE/CSEHarvardMLAStandardVancouverShortLong
PDF
Download as HTML
Select render style:
APAAuthorCBE/CSEHarvardMLAStandardVancouverShortLong
HTML
Download as Word
Select render style:
APAAuthorCBE/CSEHarvardMLAStandardVancouverShortLong
Word

ID: 6448050