Enzymatic Determination of Tissue Dihydrofolate

Abstract
A highly specific and sensitive enzymatic assay for dihydrofolate was developed. The method involves enzymatic conversion of dihydrofolate [from mouse liver] to methylenetetrahydrofolate with subsequent stoichiometric entrapment of this folate cofactor into a covalent, ternary complex with thymidylate synthase and tritiated-fluorodeoxyuridylate. The detection limit in tissue extracts was 0.25 pmol and reference dihydrofolate was quantitatively recovered in the range 0.25-2.00 pmol by this procedure.

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