Vielh P, Castellazzi M
J Immunol Methods. 1984 Oct 26;73(2):313-20. doi: 10.1016/0022-1759(84)90406-x.
We describe here an easy, sensitive and quantitative assay for the determination of adenosine deaminase activity in small populations of lymphoid cells. Determinations of enzyme activity and protein concentration are based on colorimetric reactions. All steps are directly performed in 96-well flat-bottom microtiter plates: growth of lymphoid cells (1-2 X 10(5) cells needed), detergent-mediated lysis, incubations for enzymatic assay, and reading of colorimetric reactions with an automatic multiscan spectrophotometer. Thus, screening of several hundred independent cell cultures is easily done within a day. Applications to the detection of rare adenosine deaminase variants from large cell populations and to routine medical assays are considered.
我们在此描述一种简便、灵敏且定量的测定小群体淋巴细胞中腺苷脱氨酶活性的方法。酶活性和蛋白质浓度的测定基于比色反应。所有步骤均直接在96孔平底微量滴定板中进行：淋巴细胞的培养（需要1 - 2×10⁵个细胞）、去污剂介导的细胞裂解、酶促测定的孵育以及使用自动多扫描分光光度计读取比色反应。因此，一天内轻松完成数百个独立细胞培养物的筛选。还考虑了该方法在从大量细胞群体中检测罕见腺苷脱氨酶变体以及常规医学检测中的应用。
