红细胞的毛细管区带电泳

系统研究了毛细管区带电泳对红细胞测定的操作条件和方法特点。对人、鸡、兔和猪的红血球进行了分析,结果与经典方法无异。在半小时内可一次测定10⁵个细胞,简便、精确。保留值的变异系数小于2％。实验中发现了间歇时间现象并予解释。     

毛细管电泳的最新进展

毛细管电泳是近年发展最快的分离分析技术之一。它具有高灵敏度、高分辨率、高速度等优点,广泛应用于各个领域。随着毛细管电泳技术的不断发展,逐渐出现了非水毛细管电泳,毛细管阵列电泳,毛细管电泳免疫分析,毛细管电色谱手性拆分等分支。     

毛细管电泳测定蛋白激酶A活力的新方法

建立了以毛细管电泳为基础的蛋白激酶Ａ活力测定的新方法,对其他一些激酶的活力测定具有一定的通用性。此方法是基于蛋白激酶Ａ的检测底物及其磷酸化产物容易在毛细管电泳中分开,并且可以通过在线检测进行积分定量。同时发展了连续进样技术,使能在一个电泳过程中分析十个以上的样品,大大节省分析时间和费用。     

毛细管电泳芯片及其应用

毛细管电泳芯片是近十年发展起来的一项新技术。与普通毛细管电泳相比,毛细管电泳芯片具有体积小、分离速度快、效果好、所用样品量少等优点。章介绍了毛细管电泳芯片的芯片结构及其应用等的研究进展。     

平板通道毛细管电泳

平板通道毛细管电泳又称微芯片毛细管电泳,是一门新兴的分析技术,具有小型化、自动化、快速、高效等优点。本文简要介绍了平板制作、进样、电泳分离、检测及其在生物方面的应用等。     

毛细管电泳在基因研究中的应用

讨论用于基因研究的各种方法,并指出毛细管电泳技术的优点。评述毛细管电泳(CE)用于基因分析的原理,详细介绍毛细管电泳在基因研究中的应用新进展。展望CE及其在基因研究的应用前景。     

一种新的测定蛋白激酶活性的方法：毛细管电泳测定蛋…

建立了以毛细管电泳为基础的测定蛋白激酶Ａ活性的新方法,可作为激酶测活的通用方法。此法基于底物及其磷酸化产物很容易在毛细管电泳中分开,且酶活力可用积分值计算。同时又发发展了连续井样技术,能在一次电泳中同时进行１０个以上的酶活性测定。新方法操作简单,灵敏度和精确性均优于常规的同位素法。     

平板通道毛细管电泳

平板通道毛细管电泳又称微芯片毛细管电泳,是一门新兴的分析技术,具有小型化、自动化、快速、高效等优点。本文简要介绍了平板制作、进样、电泳分离、检测及其在生物方面的应用等。     

鸭血超氧化物歧化酶(SOD)的纯化及性质研究

超氧化物歧化酶(E、C、1.15.1.1)是催化超氧阴离子起歧化反应的金属酶类,血红细胞中的SOD属Cu Zn—SOD。本文详细报道了从鸭血分离纯化SOD的改进方法(同时用猪血作对比研究)。设计了热变性、(NH_4)_2SO_4分级监析、低浓乙醇—氯仿短时去除残留血红蛋白、凝胶层析四步纯化方案,获得高产率电泳纯SOD。用紫外扫描,PAGE电泳后考马斯亮蓝和SOD活性杂色,SDS—电泳,HPLC分离和各电泳带组分的N—端测定等技术检测纯度,并进行性质研究。证明所得SOD是均一的;N—端为Ala;分子量和亚分子量各为32,359和16,500dt;并与猪血SOD对比研究其某些性质;对HPLC出现的多个活力峰及PAGE电泳显现的多条活性带进行了讨论。     

血红细胞超氧化物歧化酶(SOD)制备工艺的研究初报

本文报告了本实验室设计的由血红细胞自溶液60℃热变性, 乙醇——氯仿法除血红蛋白,旋转蒸发法减压浓缩抽去氯仿、乙醇,硫酸铵分级盐析法沉降SOD,Sepbadex G-75层析提纯SOD等步骤构成的一条成本低、设计合理、简便实用的分离纯化SOD的工艺路线。     

扬子鳄红细胞超氧物歧化酶的纯化及其性质研究

扬子鳄红细胞超氧物歧化酶的纯化及其性质研究周衍茂（安徽教育学院生物系,合肥２３００６１）尹路明（中国科学技术大学生物系,合肥２３００２６）关键词超氧物歧化酶;纯化;扬子鳄;红细胞超氧物歧化酶（的田广泛存在于各类生物组织中,是生物体内超氧自由基有效的清...     

牛红细胞超氧化物歧化酶活性中心的紫外光谱研究

应用差示分光光度法研究了牛红细胞Ｃｕ２Ｚｎ２ＳＯＤ的紫外光谱,归属和讨论了酶活性中心金属离子与配体间全部电荷转移谱带,给出了相应的配体轨道光学电负性,特别研究了涉及Ｚｎ２＋的电荷转移谱带．     

Determination of Manganese Superoxide Dismutase Activity By Direct Spectrophotometry

A method to determine Mn-superoxide dismutase activity by measuring directly the rate of decay of O₂^- in a spectrophotometer, is described. Decay of O₂^- generated by KO₂ at pH 9.5, was monitored as the fall in absorbance (A_250nm-A_360nm). Mn-superoxide dismutase was determined as the activity of cyanide-resistant superoxide dismutase, calculated from the rate of O₂^- dismutation. Mn-superoxide dismutase could be determined in the presence of a 700 times higher Cu, Zn-superoxide dismutase activity. The alkaline pH did not cause analytical problems. The assay was used to measure both Mn- and Cu, Zn-superoxide dismutase activity in mitochondrial preparations. The assay had a detection limit of 2.8 ng/ml when Mn-superoxide dismutase from E. coli was used, and the between-day CV was 5.8%. The assay is an alternative to indirect methods for detecting superoxide dismutase activity.     

The Zn-Site in Bovine Copper,Zinc Superoxide Dismutase Studied by 111Cd Pac

The active site in bovine copper, zinc superoxide dismutase (Cu₂. Zn₂ SOD) has been studied by ¹¹¹Cd time differential Perturbed Angular Correlation (PAC) on enzyme with Zn²⁺ replaced by excited 'Cd²⁺. The PAC spectra obtained for both the oxidized and the reduced form of Cu₂Cd₂SOD show no asymmetry between the two Zn-sites in the dimeric enzyme. The spectv further reveal that a significant change has taken place at the Zn-site in the reduced form compared to the oxidized form.

Semi-empirical calculations based on the Angular Overlap Model (AOM) and coordinates from the crystal structure of the native enzyme are in agreement with the experimental PAC data of the oxidized enzyme. The results indicate that Cd²⁺ coordinates in the same manner as Zn²⁺ and that the crystal structure of SOD is valid for the enzyme in solution. The PAC spectrum of the reduced enzyme can be explained by extending the AOM calculations to the enzyme in the reduced form and assuming that the imidazol ring of His61 is no longer bridging the copper and cadmium ions in the reduced state.     

Mechanism and Atomic Structure of Superoxide Dismutase

The active site Cu ion in Cu,Zn superoxide dismutase is alternately oxidized and reduced during the enzymatic dismutation of superoxide to hydrogen peroxide and molecular oxygen. For oxidized Cu,Zn superoxide dismutase, an atomic structure has been determined for the human enzyme at 2.5 A resolution. The resolution of the bovine enzyme structure has been extended to 1.8 A. Atomic resolution data has been, collected for reduced and inhibitor-bound Cu,Zn superoxide dismutases. and the interpretation of the' electron density difference maps is in progress. The geometry and molecular surfaces of the active sites in these structures, together with biochemical data, suggest a specific model for the enzyme mechanism. Similarities in the active site geometry of the Mn and Fe superoxide dismutases with the Cu.Zn enzyme suggest that dismutation in these enzymes may follow a similar mechanism.     

超氧化物歧化酶在氧自由基所引起的癌变中的作用

超氧化物歧化酶是一类抗氧化酶,它能催化超氧阴离子自由基的歧化反应,对机体起保护作用。氧自由基在某些情况下会对机体产生损伤作用。文章就氧自由基的产生、氧自由基与致癌的关系,以及超氧化物歧化酶在氧自由基所引起的癌变中的作用等方面进行了综述。     

中药合剂CB在晶状体白内障形成中对巯基及二硫键的作用

用白内障诱发剂三硝基甲苯、平阳霉素、亚硒酸钠和半乳糖分别加入大鼠晶状体培养基中,共同培养２４ｈ,同时在各培养基中分别加入中药合剂ＣＢ,以观察其药效,用维生素Ｃ作对照。结果表明中药合剂ＣＢ能够保护非蛋白质巯基免致氧化,抑制蛋白质巯基交联,降低晶状体不溶性蛋白质中二硫键含量,故中药合剂ＣＢ有可能作为抗白内障药物应用于临床。     

嗜热脂肪芽孢杆菌DNA解链蛋白BstH2的纯化及性质研究

以嗜热脂肪芽孢杆菌为材料,通过ＰｏｌｙｍｉｎＰ沉淀、硫酸铵分级及ＤＥＡＥ纤维素、磷酸纤维素、Ｂｌｕｅ－Ｓｅｐｈａｒｏｓｅ、ＦＰＬＣＭｏｎｏＱ、ＦＰＬＣ Ｓｕｐｅｒｏｓｅ１２等柱层析,得到了部分纯化ＤＮＡ解链蛋白ＢｓｔＨ２。ＢｓｔＨ２具有受ＤＮＡ促进的ＡＴＰ酶活力,没类型的核酸对ＢｓｔＨ２的ＡＴＰ酶活力的促进作用没。ＢｓｔＨ２在５５℃有最高ＡＴＰ酶活力。这种活力受大肠杆菌单链ＤＮＡ结合蛋白的抑制及随