Isolation of a complex of respiratory burst oxidase components from resting neutrophil cytosol.
Isolation of a complex of respiratory burst oxidase components from resting neutrophil cytosol.
复制标题
从静息中性粒细胞胞质中分离呼吸爆发氧化酶成分复合物。
DOI：
10.1021/bi00176a021
复制
发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
Babior,BM
中科院分区：
文献类型：
--
作者：
Park,JW;Benna,JE;Scott,KE;Christensen,BL;Chanock,SJ;Babior,BM
材料与方法中性粒细胞组分的制备。中性粒细胞胞浆和质膜的制备如前所述(Park等人，1992)。简而言之，通过葡聚糖沉淀和新鲜提取的柠檬酸盐抗凝血液的Ficoll-Hypaque分级，从正常受试者中获得中性粒细胞。中性粒细胞以10×细胞/毫升的浓度悬浮在改进的松弛缓冲液(100 mM KC1/3.0 mM Na C l/3.5 mM MgCl2/10 mm管状缓冲液，pH 7.3)中，通过氮气空化和Percoll离心法按Borregaard等人的方法制备质膜和胞浆(Borregaard等，1983)。将细胞液和细胞膜分成等份，在-70℃保存直至使用。
MATERIALS AND METHODSPreparation of Neutrophil Fractions. Neutrophil cytosol and plasma membrane were prepared as described previously (Park et al., 1992). Briefly, neutrophils were obtained from normal subjects by dextran sedimentation and Ficoll-Hypaque fractionation of freshly drawn citrate-anticoagulated blood. The neutrophils were suspended at a concentration of 10* cells/mL in a modified relaxation buffer (100 mM KC1/3.0 mM NaCl/3.5 mM MgCl2/10 mM PIPES buffer, pH 7.3), and plasma membrane and cytosol were prepared by nitrogen cavitation and centrifugation through Percoll by the method of Borregaard (Borregaard et al., 1983). Both cytosol and membrane were divided into aliquots and stored at-70 C until use.