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affinity column     
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  亲和柱
     Construct a prokaryotic expression vector pET-28a-LG1 by DNA recombination technique and express hLNα4LG1 fusion protein in BL21(DE3)/pET system. The expressed product was identified by SDS-PAGE,purified by Ni-NTA affinity column and analyzed by Western blot.
     用DNA重组技术构建原核表达载体pET28aLG1,用BL21(DE3)pET系统表达hLNα4LG1融合蛋白,SDSPAGE鉴定表达产物,用NiNTA亲和柱对表达蛋白进行纯化,并进行Westernblot分析。
短句来源
     Fluorimetric Determination of Aflatoxin B_1 in Peanut and Corn With Monoclonal Antibody Affinity Column Cleanup and Bromine Treatment
     单克隆抗体亲和柱分离荧光法测定花生和玉米中黄曲霉毒素B_1的研究
短句来源
     About 15-20 mg of recombinant protein was purified from 1 L of culture supernatant of recombinant E. coli by Ni~2+-NTA affinity column chromatography,and reached a purity of more than 80%.
     通过Ni2+-NTA亲和柱,从1L诱导产物中纯化出约1520mg重组蛋白,纯度在80%以上。
短句来源
     The isolating effect of receptor affinity chromatogragy for CT-B or CT are 71.6% or 48.3% respectively. IgG affinity column for CT-B or CT are 50% or 51.7% respectively.
     受体亲和柱对CT-B和CT的分离效率分别为71.6%或48.3%,抗体亲和柱分别为50%或51.7%。
短句来源
     Silica gel (for chromatography) and agarose 6B were used as carrier for chromatography, then Cu 2+ was chelated to form metal chelate affinity column. The recombinant human Cu,Zn superoxide dismutase (rh Cu,Zn SOD) was purified in one step metal chelate affinity chromatography by Cu 2+ IDA silica gel column and Cu 2+ IDA Agarose 6B column.
     以层析硅胶Agarose 6B ,制备了Cu2 + 螯合亲和载体 ,利用合成的Cu2 + IDA 硅胶亲和柱和Cu2 + IDA Agarose 6B亲和柱分离纯化了重组人Cu ,Zn SOD ,并比较了这两 2亲和载体的纯化效果和性能。
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  亲和层析柱
     An affinity column based on 5'-AMP-QT_4 was applied to purify lactate dehydrogenase (LDH).
     进行了5′—AMP—QT_4亲和层析柱纯化乳酸脱氢酶(LDH)的试验。
短句来源
     By using 5 G 1-sepharose 4 B immune affinity column, we recovered approximately 160 μg purified and soluble IL-2 receptor (sIL-2 R ) from 500ml PHA-stimulated human PBL supernatant.
     利用溴化氰活化Sepharose 4B制备了免疫吸附亲和层析柱5G_1-Sepharose 4B,从500ml PHA刺激的PBL培养上清中纯化了sIL-2R约160μg。
短句来源
     The purities of the recombinant proteins achieve 85% on glutathione-sepharose 4B affinity column.
     经谷胱甘肽-sepharose 4B亲和层析柱一步纯化后,4种蛋白的纯度均达到85%以上。
短句来源
     Methods Purity and yield ratio and conjugated activity of purified Fab fragment were analyzed with three purified ways of goat anti human Fab affinity chromatography and 14F7 monoclonal antibody affinity column, as well as Ion exchange Size exclusion column.
     方法采用羊抗人Fab抗体亲和层析柱、14F7单克隆抗体亲和层析柱、离子交换分子筛层析柱,分别纯化由酵母工程菌(GS115/Fab)发酵的重组人抗HBsAgFab抗体,并对3种纯化方法所得Fab抗体的纯度、收率、与HBsAg的结合活性进行比较。
短句来源
     Circumsporozoite protein (CSP) ,with molecule weight of 4oKd, was purified from sporozoite extract through MS1-Sepharose 4B affinity column.
     用MS1-Sepharose4B亲和层析柱能自子孢子可溶性蛋白中提取CSP。
短句来源
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  亲和层析
     An affinity column based on 5'-AMP-QT_4 was applied to purify lactate dehydrogenase (LDH).
     进行了5′—AMP—QT_4亲和层析柱纯化乳酸脱氢酶(LDH)的试验。
短句来源
     The fusion protein was eluted from affinity column that is about 85Kd, 58Kd rhOPN and 26Kd GST are obtained after cleavage.
     亲和层析的洗脱溶液中是分子量为85 Kd的融合蛋白GST-OPN,该蛋白裂解为分子量58 Kd的rhOPN和26 Kd的GST。
短句来源
     By using 5 G 1-sepharose 4 B immune affinity column, we recovered approximately 160 μg purified and soluble IL-2 receptor (sIL-2 R ) from 500ml PHA-stimulated human PBL supernatant.
     利用溴化氰活化Sepharose 4B制备了免疫吸附亲和层析柱5G_1-Sepharose 4B,从500ml PHA刺激的PBL培养上清中纯化了sIL-2R约160μg。
短句来源
     The purities of the recombinant proteins achieve 85% on glutathione-sepharose 4B affinity column.
     经谷胱甘肽-sepharose 4B亲和层析柱一步纯化后,4种蛋白的纯度均达到85%以上。
短句来源
     Methods Purity and yield ratio and conjugated activity of purified Fab fragment were analyzed with three purified ways of goat anti human Fab affinity chromatography and 14F7 monoclonal antibody affinity column, as well as Ion exchange Size exclusion column.
     方法采用羊抗人Fab抗体亲和层析柱、14F7单克隆抗体亲和层析柱、离子交换分子筛层析柱,分别纯化由酵母工程菌(GS115/Fab)发酵的重组人抗HBsAgFab抗体,并对3种纯化方法所得Fab抗体的纯度、收率、与HBsAg的结合活性进行比较。
短句来源
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  亲和色谱柱
     Preparation of a Concanavalin A Immobilized Affinity Column and Its Application in the Structural Analysis of Ribonuclease B
     伴刀豆球蛋白亲和色谱柱的制备及其在糖蛋白核糖核酸酶B结构分析中的应用
短句来源
     MethodsAfter binding a monoclonal antibody against PF4 (SZ 95) to CNBr activated sepharose 4B, a SZ 95 sepharose 4B affinity column was got. Proteins, which were purified from triton X 100 solubilized platelet solution by SZ 95 sepharose 4B affinity chromatography, were characterized by 15% SDS PAGE and Dot blot.
     方法将单克隆抗体SZ 95 IgG与溴化氰活化的Sepharose 4B凝胶连接成亲和色谱柱SZ 95 IgG Sepharose 4B ,人血小板破碎液经此亲和色谱柱上样后 ,经洗脱获得PF4 ,采用 15 %SDS 聚丙烯酰胺凝胶电泳鉴定其纯度 ,用点印迹鉴定其免疫活性。
短句来源
     ResultsThe specific inhibition activity of elution peak of ACEI affinity column was up to 1 450 U/mg, and the recovery rate of activity was 12.1%. The purity of eluent was 906 times higher than that of the crude extract.
     结果亲和色谱柱洗脱液峰值管比抑制活力达 14 5 0u/mg,为粗提液的 90 6倍 ,活力回收率为 12 .1%。
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      affinity column
    The polyclonal antibodies were tentatively purified and then used as ligands of an affinity column.
          
    Pallas crude venom was fractionated by this affinity column and then by Mono Q on fast protein liquid chromatography (FPLC).
          
    In order to separate and purify some of the enzymes contained in porcine pancreas, operation of an ion-exchange column in series with an affinity column has been used.
          
    By use of frontal analysis on an affinity column we have examined the binding interaction of berberine chloride (BC), a major active constituent of coptis, with bovine serum albumin (BSA) in 40?mM phosphate buffer, pH?7.0.
          
    Total water soluble protein (WSP) was 133 ± 68 (SD) mg/100 g (6) cuticle, whereas affinity column bound protein, CA, was 191 ± 108 (SD) μg/100 g (6) cuticle.
          
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    The affinity column purified anti-DNP antibodies from secondary response of a single Wistar rat were used to immunize a rabbit. The serum of which was first absorbed with immunoglobulins of normal Wistar rats and then precipitate with ammonium sulphate. The anti-idiotypic specificities of the antibodies thus obtained were identified with ELISA, in which the walls of plate were coated with affinity column purified anti-DNP antibodies. With self Ig separated from preimmunized serum as control, the...

    The affinity column purified anti-DNP antibodies from secondary response of a single Wistar rat were used to immunize a rabbit. The serum of which was first absorbed with immunoglobulins of normal Wistar rats and then precipitate with ammonium sulphate. The anti-idiotypic specificities of the antibodies thus obtained were identified with ELISA, in which the walls of plate were coated with affinity column purified anti-DNP antibodies. With self Ig separated from preimmunized serum as control, the results showed that the antibodies were directed at the idiotypic determinants on anti-DNP antibodies of the Wistar rat. In order to test for cross reactive idiotypes on anti-DNP antibodies from other Wister rats, a reverse ELISA was used, i. e. the above characterized antiidiotypic antibodies rather than the anti-DNP antibodies were used to coat the plate. It was shown that the reverse ELISA could specifically detect and semiquantatively determine the idiotypes directly from serum. This reverse ELISA could afford facilities for researches in idiotype expression. The effect of blocking DNP combining site on the idiotype-antiidiotype reactions was also studied.

    本文以“逆向”ELISA 测定 Wistar 大鼠抗 DNP抗血清中抗体的独特型,即以抗独特型抗体为涂布物来测定抗血清的效价。实验结果表明“逆向”ELISA可以专一地半定量测定大鼠抗 DNP抗血清的独特型,为进一步研究抗独特型抗体对抗体独特型表达的调节作用打下基础。同时,DNP-BSA封闭大鼠抗 DNP抗体的实验初步表明,这抗体的独特型决定簇位于抗体分子的抗原结合部位,至少十分接近抗原结合部位。

    We report here the serological properties of McAbs secreted by the hybrids of 8 cell lines in which 4 were anti-whole IgG, 3 were anti-IgG with no response against IgG4 and 1 was anti-IgG except IgG3. There appeared to be some differences in the effective titers and antigen-binding capacities between each of these McAbs observed. McAbs A2 and B1 were selected for the combination according to the combining test, and they were subsequently used to detect the IgG amounts of normal individuals. The results correlated...

    We report here the serological properties of McAbs secreted by the hybrids of 8 cell lines in which 4 were anti-whole IgG, 3 were anti-IgG with no response against IgG4 and 1 was anti-IgG except IgG3. There appeared to be some differences in the effective titers and antigen-binding capacities between each of these McAbs observed. McAbs A2 and B1 were selected for the combination according to the combining test, and they were subsequently used to detect the IgG amounts of normal individuals. The results correlated well with that of conventional antisera (r= 0.90). McAbs were utilized to prepare the affinity column for chromatography. The yield of purified human IgG by chromatography was 7.3 times higher than that obtained with conventional antibodies as affinity reagent. It is considered from above observation that the investigation and preparation of such McAbs may provide effective reagents for clinical immunological determinations.

    本文报道8株杂交瘤细胞分泌的单克隆抗体的血清学特性,其中4种属抗全IgG,3种是对IgG_4无反应的抗IgG,1种是对IgG_3无反应的抗IgG。各单克隆抗体之间的效价和结合抗原的能力有所差异。根据组合试验,选出A_2与B_1二种单克隆抗体进行组合,用以检测正常人血清中IgG含量,与常规抗血清相比较呈良好的相关性(r=0.90)。用以制备亲和层析柱,纯化人IgG,得率比常规抗血清高7.3倍。据此认为,研制这类单克隆抗体将为临床免疫检测提供有效的试剂。

    The nitrate reductase preparation from rice leaves was purified by affinity chromatography. Two peaks of NADH: nitrate reductase activity were found when the enzymes was eluted from Blue Dextran-Sepharose 4B affinity column by 100μM NADH and 0.25 M KNOa solution successively. These two fractions of nitrate reductase were designated as NADH- and KNO_3-eluted nitrate reductase respectively. This suggested that nitrate reductase was bound to the affinity column through two binding sites: One was a NADH-binding...

    The nitrate reductase preparation from rice leaves was purified by affinity chromatography. Two peaks of NADH: nitrate reductase activity were found when the enzymes was eluted from Blue Dextran-Sepharose 4B affinity column by 100μM NADH and 0.25 M KNOa solution successively. These two fractions of nitrate reductase were designated as NADH- and KNO_3-eluted nitrate reductase respectively. This suggested that nitrate reductase was bound to the affinity column through two binding sites: One was a NADH-binding site, and the other was a NO_3-binding site. In eluting nitrate reductase from the affinity column, the nitrate may act either by increasing the ionic strength or through a substrate effect, or both. When KNO_3-eluted nitrate reductase was collected and again passed through the affinity column for the second or the third time, there were still two peaks of nitrate reductase, but the ratio of the activity of KNO_3-eluted nitrate reductase to that of NADH-eluted one decreased. After the third time of affinity chromatography, very little KNO_3-eluted nitrate reductase activity remained. However, by means of 7.5% polyacrylamide gel and SDS-polyacrylamide gel electrophoresis, both NADH- and KNO_3-eluted nitrate reductase showed the same electrophoresis pattern.

    水稻叶片NR在Blue Dextran-Sepharose 4 B亲和层析法的纯化过程中,继100μMNADH洗脱出现一个酶活峰后,再用0.25MKNO_3洗脱又出现一个更高的酶活峰。酶与BlueDextran-Sepharose 4B有两个结合位点,它们可能就是酶与底物NADH和NO_3的结合位点。NADH能和Blue Dextran竞争与酶结合而把NR从亲和柱上洗脱下来。硝酸盐除了具一般盐类的离子强度外,还有明显的底物效应。如果将KNO_3-洗脱的NR再反复进行亲和层析,又可得到NADH-洗脱和KNO_3-洗脱的两部分NR。这两部分NR在聚丙烯酰胺凝胶电泳和SDS-聚丙烯酰胺凝胶电泳上都呈现一条相同的蛋白带。 水稻NR的全酶分子量约330Kd,亚基分子量约57Kd,故全酶可能由6个相同亚基组成,酶活的pH范围为6.5~8.5,最适pH为7.5;酶对底物NO_3-和NADH的K_m值分别为3.3×10~(-4)M和2.9×10~(-5)M。

     
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