DBPDADivision of Budget Policy and Data Analysis
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In step 3, HZS combines with DBPDA to form a compound that is fluorescent and has ultraviolet absorbance.
[1] Nonstandard abbreviations: PLP, pyridoxal 5'-phosphate; tHCY, total homocysteine; REA, radioenzymafic assay; rHCYase, recombinant homocysteine [alpha],[gamma]-lyase; and DBPDA, N,N-dibutylphenylenediamine.
In reaction II, the [H.sub.2]S combines with DBPDA to form 3,7-bis(dibutyl amino)phenothiazine-5-ium chloride, which is highly fluorescent.
The enzymatic reaction was stopped by the addition of 50 [micro]L of chromophore reagent (20 mmol/L DBPDA in 3 mol/L [H.sub.2]S[O.sub.4]), followed by 30 [micro]L of oxidizing agent [5 mmol/L potassium ferricyanide in 10 mmol/L sodium phosphate buffer (pH 7.2)].