Chromogenic substrate with a pH indicator dye
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United States of America Patent
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Issued Date -
Nov 4, 2004
app pub date -
Apr 29, 2003
filing date -
Apr 29, 2003
priority date (Note) -
Abandoned
status (Latency Note)
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Abstract
A composition for an enzyme-linked assay is described which comprises a chromogenic substrate that produces a detectable color with a maximum absorbance at one wavelength in reactions of the assay which contain the enzyme and a pH indicator dye that produces a detectable color with a maximum absorbance at a second wavelength when the pH of the reactions of the assay is changed to terminate the reactions. The chromogenic substrate enables colorimetric detection of the bindable substance in positive reactions of the assay and the pH indicator dye enables negative reactions of the assay to be calorimetrically distinguished from false negative reactions. Preferably, the composition is used in enzyme-linked immunoassays such as ELISAs. Most preferably, the enzyme is a peroxidase. In a further preferred embodiment, the chromogenic substrate is 3,3',5,5'-tetramethylbenzidine (TMB) and the pH indicator dye is preferably selected from the group consisting of cresol red and m-cresol purple.
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