Determination of arginase activity in macrophages: a micromethod

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Abstract

We propose a modification of Schimke's method for urea determination as a valuable micromethod for measuring arginase in activated macrophages. The method exhibits the following advantages: (a) it uses small amounts of samples (approximately 25 000 macrophages per assay); (b) it does not interfere with other related metabolites that are also present in the activated macrophage such as citrulline or arginine; (c) saturating concentrations of the substrate arginine can be used; and (d) it is much more sensitive than Schimke's method and can detect small amounts of urea, in the order of 0.02 μmol.

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    Citation Excerpt :

    Then, absorbance was noted at 550 nm. The assay calculation was done with a standard curve made by using 2-fold serial dilutions of 200 mM urea (200–3.12 mM) (Corraliza et al., 1994). The arginase enzyme activity was expressed by µM urea/mg protein.

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