Kits for assaying enzyme-mediated oxidative demethylation

Abstract

Methods of assaying enzyme-mediated oxidative demethylation are provided according to embodiments of the present invention which includes combining, under reaction conditions, an oxidative demethylation enzyme, a substrate for the oxidative demethylation enzyme and a formaldehyde detection reagent. Detection of fluorescence is indicative of formaldehyde generated by oxidative demethylation of the substrate by the enzyme, the fluorescence resulting from reaction of formaldehyde and the formaldehyde detection reagent.

1 . A kit comprising:
 4-amino-3-penten-2-one; and   at least one reagent for use in a method of assaying formaldehyde generated by oxidative demethylation of a substrate by an oxidative demethylation enzyme.   
     
     
         2 . The kit of  claim 1 , wherein the reagent is an oxidative demethylation enzyme. 
     
     
         3 . The kit of  claim 1 , wherein the reagent is a substrate for an oxidative demethylation enzyme. 
     
     
         4 . The kit of  claim 1 , wherein the reagent is an oxidative demethylation enzyme cofactor. 
     
     
         5 . The kit of  claim 4 , wherein the oxidative demethylation enzyme cofactor is selected from the group consisting of Fe(II), alpha-ketoglutarate, FAD, NADH, NADPH and ascorbic acid. 
     
     
         6 . The kit of  claim 1 , wherein the reagent is a co-enzyme for use in a method of assaying formaldehyde generated by oxidative demethylation of a substrate by an oxidative demethylation enzyme. 
     
     
         7 . The kit of  claim 1 , wherein the reagent is a reaction buffer. 
     
     
         8 . The kit of  claim 1 , wherein the reagent is a cell lysis buffer. 
     
     
         9 . The kit of  claim 1 , further comprising a reaction vessel for performing oxidative demethylation of a substrate by an oxidative demethylation enzyme. 
     
     
         10 . The kit of  claim 1 , further comprising a control or standard selected from the group consisting of: a positive control reagent, a negative control reagent and a standard. 
     
     
         11 . The kit of  claim 1 , further comprising a positive control sample, the positive control sample comprising formaldehyde.