I am trying to prepare Ellman's reagent solution to detect Glutathione. Before mixing with my sample. The solution already has slight yellow color. Is that correct or I already contaminated it?
The following is the recipe I used for the Ellman's reagent solution.
1 mL of 0.4 M Tris–HCl, pH 8.9 containing 20mM EDTA and 25mL of 10mM DTNB
Cho, A. K.; Sioutas, C.; Miguel, A. H.; Kumagai, Y.; Schmitz, D. A.; Singh, M.; Eiguren-Fernandez, A.; Froines, J. R. Redox Activity of Airborne Particulate Matter at Different Sites in the Los Angeles Basin. Environmental Research. 2005, 99, 40–47.
A solution of this compound produces a measurable yellow-colored product when it reacts with sulfhydryls. Consequently, Ellman’s Reagent is very useful as a sulfhydryl assay reagent because of its specificity for -SH groups at neutral pH, high molar
extinction coefficient and short reaction time.
Color is shown when the DTNB solution is mixed with the buffer before mixed with the sample. Is that correct?
Dear Nai-Yuan Liu , cited below is an article which is available on RG: In this paper the color of DTNB (5,5′-dithiobis(2-nitrobenzoic acid)) has been described as pale yellow.
Article Tissue Sulfydryl Groups
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