全染料染色法测定末端脱氧核苷酰转移酶活力

Stains-all staining Assay for Terminal De-oxynucleotidyl Transferase Activity.

介绍一种末端脱氧核苷酰转移酶活力测定的新方法，利用全染料（ｓｔａｉｎｓ─ａｌｌ）对单链寡聚核苷酸片段灵敏度较高的特点，建立了电泳─全染料染色法测定ＴｄＴ酶活力，ＴｄＴ酶促加长的寡聚核苷酸片段样本，经聚丙烯酰胺凝胶电泳分离、全染料染色后即可根据寡聚核苷酸片段的泳动距离推算ＴｄＴ酶活力。此法所用试剂便宜，操作简便，无需大型仪器及灵敏度较高，适用于临床上白血病患者白细胞中ＴｄＴ酶活力的半定量检测及基因工程中ＴｄＴ工具酶活力的测定。

polyacrylamide gel electrophoresis-stains-allstaining assay for terminal deoxynucleotidyltransferase (TdT) activity in purified TdT andleucocytic homogenate of normal and leukemicpatients has been developed. The results showthat stains-all[(1-ethy1=2-[3-(1-ethyl-naphtho[1.2-d] thiazolin-2 ylidene)-2-methylpropenyl]naphtho [1,2-d]-thiazoliumbromide] is highly specific for single strand oli-go-deoxynucleotidyl fragment; no other cellcomponent produces influence.The methodpermits the estimation of TdT activity as lowas 3U.It is a simple assay suitable for clinicalanalysis of TdT in human leukemias and provd-ing information useful in classifying haemao-logical neoplasms.