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组织化学和细胞组分研究表明,碱性磷酸酶(APase)存在于B细胞性白血病和淋巴瘤以及淋巴母细胞样B细胞株的细胞中,但T淋巴瘤细胞株或骨髓样细胞中则缺如。本文报道建立一种简单、快速检测小鼠脾脏细胞膜APase活性的微量滴定试验。将正常或培养的细胞(4×10~5,悬浮于50μ1PBS)加入聚苯乙烯微量滴定板小孔内,经离心后用0.25M戊二醛使贴壁细胞固定,继用含0.1%吐温的PBS洗涤后,加100μl含1mg/ml对硝基苯磷酸盐(底物)和10~(-3)M MgCl_2的0.05M碳酸钠缓冲液(pH9.8),在室温下放置30分钟,随即加入50μ10.3M NaOH终止反应。用平板多道扫描读数仪测量各孔在405 um的消光度。用本法检测了在体外经LPS和PWM激活的小鼠脾细胞质膜的
Histochemistry and cellular components studies have shown that alkaline phosphatase (APase) is present in cells of B-cell leukemia and lymphoma and lymphoblastoid B cell lines, but lacking in T-lymphoma cell lines or bone marrow-like cells Such as. This article reports the establishment of a simple and rapid detection of mouse spleen cell membrane APase activity microtiter test. The normal or cultured cells (4 × 10 ~ 5, suspended in 50μ1 PBS) was added to polystyrene microtiter plate wells, after centrifugation with 0.25M glutaraldehyde adherent cells fixed, followed by containing 0.1% Tween After washing with PBS, 100 μl of 0.05 M sodium carbonate buffer (pH 9.8) containing 1 mg / ml of p-nitrophenylphosphate (substrate) and 10 -3 M MgCl 2 was added and left to stand at room temperature for 30 minutes, The reaction was then quenched by the addition of 50 [mu] 10.3 M NaOH. The flatness multichannel scanning reader was used to measure the extinction of each well at 405 um. This method was used to examine the plasma membrane of mouse splenocytes activated by LPS and PWM in vitro