Van Laarhoven J P, Spierenburg G T, De Bruyn C H
J Immunol Methods. 1980;39(1-2):47-58. doi: 10.1016/0022-1759(80)90293-8.
A methodology is presented for systemic analysis of purine enzymes in small lymphocyte subfractions. For the determination of 7 different enzymes of purine metabolism *hypoxanthine-guanine phosphoribosyltransferase (HG-PRT), adenine phosphoribosyltransferase (A-PRT), adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), adenosine kinase (AK), 5'-nucleotidase (5'N), and AMP-deaminase) less than 200,000 peripheral blood lymphocytes are needed. 1000-6000 lyophilised lymphocytes are incubated in micro-incubation vessels (3 microliter) with radioactive substrates for 15-180 min. Separation of substrates and products is achieved by thin-layer chromatography on PEI-cellulose. Addition of BSA to the incubation mixtures results in higher specific enzyme activities and narrower ranges of mean values of a control group.
本文介绍了一种用于小淋巴细胞亚群中嘌呤酶系统分析的方法。为了测定嘌呤代谢的7种不同酶（次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶（HG-PRT）、腺嘌呤磷酸核糖转移酶（A-PRT）、腺苷脱氨酶（ADA）、嘌呤核苷磷酸化酶（PNP）、腺苷激酶（AK）、5'-核苷酸酶（5'N）和AMP脱氨酶），需要少于200,000个外周血淋巴细胞。将1000 - 6000个冻干淋巴细胞在微量孵育容器（3微升）中与放射性底物孵育15 - 180分钟。通过在聚乙烯亚胺 - 纤维素上进行薄层层析来分离底物和产物。在孵育混合物中添加牛血清白蛋白（BSA）可导致更高的比酶活性以及对照组平均值范围更窄。
