A sensitive radiochemical assay for adenosine deaminase

Adenosine deaminase (EC 3.5.4.4) (ADA) is assayed by incubating [8- 14C]adenosine with the enzyme containing solution for 2 hr at pH 7.4 and 37°C. The trichloroacetic acid protein free filtrate is acid hydrolyzed and separated on a 2.5 × 0.2 cm 2 Dowex-50 column where the hypoxanthine is eluted quan...

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Veröffentlicht in:	Clinical immunology and immunopathology 1976-03, Vol.5 (2), p.173-176
Hauptverfasser:	Mudawwar, Fuad, Jebara, Haifa, Geha, Raif, al-Khalidi, Usama
Format:	Artikel
Sprache:	eng
Schlagworte:	Adenine - isolation & purification Adenosine Deaminase - analysis Adenosine Deaminase - blood Humans Hypoxanthines - isolation & purification Nucleoside Deaminases - analysis Radiochemistry
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Zusammenfassung:	Adenosine deaminase (EC 3.5.4.4) (ADA) is assayed by incubating [8- 14C]adenosine with the enzyme containing solution for 2 hr at pH 7.4 and 37°C. The trichloroacetic acid protein free filtrate is acid hydrolyzed and separated on a 2.5 × 0.2 cm 2 Dowex-50 column where the hypoxanthine is eluted quantitatively in the first 15 ml of N HCl and counted. This assay is about 1000 times more sensitive than the spectrophotometric. It is linear up to 200 μIU/sample, reproducible, lends itself to serial determinations, and can be performed on turbid solutions.
ISSN:	0090-1229 1090-2341
DOI:	10.1016/0090-1229(76)90022-2