Application of a fluorogenic reagent, ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate for detection of cystine-containing peptides.

Abstract:

:A simple, sensitive and reliable method for the detection of cystine-containing peptides has been developed. A peptide bridged with a disulfide bond was reduced and cleaved with tributylphosphine, and then coupled with a thiol specific reagent, ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate, under alkaline conditions. After incubation at 60 degrees C for 1 h, the fluorescent derivative formed was measured with excitation at 385 nm and emission at 515 nm. The intensity of fluorescence labeled to the peptide was very stable and the peptide containing disulfide was quantitatively determined in the range of 100 pmol to 10 nmol, when oxidized glutathione was used as a standard. This method was very useful for specific detection of cystine-containing peptides in the column effluents on reverse phase high performance liquid chromatography.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Sueyoshi T,Miyata T,Iwanaga S,Toyo'oka T,Imai K

doi

10.1093/oxfordjournals.jbchem.a135241

subject

Has Abstract

pub_date

1985-06-01 00:00:00

pages

1811-3

issue

6

eissn

0021-924X

issn

1756-2651

journal_volume

97

pub_type

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