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The aim of this study was to investigate the expression of mPGES-2 protein in mouse oviduct during the early pregnancy, pseudopregnancy, and estrous cycle by immunohistochemistry and Western blot. The regulation of mPGES-2 protein in mouse oviducts was also studied under delayed implantation, steroid hormonal treatments and oviductal ligation.

本实验主要利用早期妊娠、假孕、输卵管结扎、发情周期、激素调节以及延迟着床等实验模型,结合免疫组织化学和Western blot 的实验方法,详细研究了mPGES-2 蛋白在小鼠输卵管中的表达与调节。mPGES-2 蛋白主要定位于不同时期的伞部和壶腹部的上皮纤毛细胞中,在伞部和壶腹部肌层以及峡部和宫管连接处的上皮和肌层中未检测到mPGES-2 蛋白的表达。

Reverse-transcription PCR assays were used to detect the mRNA expression level of related regulatory genes such as p15, p16, p21, p27, p57, surviving, cyclin B, cdc2 and chk1, and Western blot assay to detect the level of protein expression and phosphated change such as survivin, cdc2 and chk1. Results:(1) K562 cell arrested on G2/M phase were obviously increased after co-cultured with 2 to 10μmol/L Arsenic trioxide for 24 hours. The ratios of control group, 2μmol/L, 5μmol/L and 10μmol/L groups were 22.6±3.4%, 27.2±2.3%, 43.8±4.5% and 36.7±4.1%, respectively.

体外培养K562细胞,以不同浓度AS_2O_3作用不同时间后采用PI染色、流式细胞仪检测药物作用后细胞的周期分布改变;逆转录酶多聚酶链扩增方法检测细胞周期相关调节基因(p15、p16、p21、p27、p57、survivin、cyclinB、cdc2、chk1)的mRNA表达变化;Western blot方法检测细胞周期相关调节蛋白表达及磷酸化改变(survivin、cdc2、cdc2-p、chk1)。

Osteoblasts were detected by alkaline phosphatese and calcium salt staining. The expression of osteopontin was measured by Western blotting.

成骨细胞鉴定采用组织化学方法行碱性磷酸酶和钙化斑块染色同时应用Western blotting法测定骨桥蛋白表达。

Detect with Western Blot protein of MAb counterpoise form and the natural phosphoric acid is enzymatic reactivity in the cell, obtain 6 individual plant to be able to be stabilized in all secrete the individual plant of cross tumour cell that fights phosphoric acid enzymatic MAb, go enzymaticly to study double particularity phosphoric acid further phosphation action mechanism and its activation formerly in mitosis way of enzymatic signal of albumen phosphoric acid turns guide provide strong tool.

用Western blot检测mAb对重组蛋白和细胞中天然磷酸酶的反应性,共获得6株可稳定分泌抗磷酸酶mAb的杂交瘤细胞株,为进一步探究双特异性磷酸酶的去磷酸化功能机理以及其在有丝分裂原激活蛋白磷酸酶信号途径中的信号转导提供强有力的工具。

Cent of 玀 HCC97H cell is contrast group of group, HGF group, brilliant mildew vegetable, reach Western Blot with RT-PCR respectively the metabolic circumstance of afore-mentioned target albumen after change of level of Sp1 of analysis and VEGF MRNA, protein expression and Sp1 level, phosphation and applied Sp1 block break element of agent brightness mildew.

将MHCC97H细胞分为对照组、HGF组、光辉霉素组,分别用RT-PCR及Western blot分析和VEGF mRNA、蛋白质表达及Sp1水平、磷酸化Sp1水平变化以及应用Sp1阻断剂光辉霉素后上述目标蛋白的变化情况。

The method uses EGCG of different pH indicator to process HT-29 cell, colorimetric of 4 Zun salt experiments (the growth of the cell after MTT) law observes processing circumstance; sheds type cell art detects cellular wither dies clean of circumstance;Western Blot detects skin grows factor accepts put oneself in another's position , cell adjusts kinase phosphation level.

方法采用不同浓度的EGCG处理HT-29细胞,四唑盐比色试验法观察处理后细胞的生长情况;流式细胞术检测细胞凋亡情况;Western Blot洁检测表皮生长因子受体、细胞外信号调节激酶磷酸化水平的表达。

The expression of SRp38 in mouse retina was investigated by Western blot and immunohistochemistry analyses. The result shows that the expression of SRp38 proteins in mouse retina is region-specific, with extensive distribution in the outer and inner plexiform layers, inner nuclear layer and ganglion cell layer, but no expression in outer nuclear layer.

利用Western blot 以及免疫组织化学方法研究了SRp38蛋白在小鼠视网膜中的表达以及分布情况,结果显示,SRp38蛋白在视网膜中的表达具有区域特异性,在外网层、内核层、内网层以及节细胞层中均有表达,而在外核层无表达。

Methods Purified 5-LO fragment was injected into male pronucli and the firtilized eggs were transplanted into pseudopregnant mice. PCR and Southern blot were used to detect the genotype of DNA separated from the newborn mouse tail tissues. RT-PCR and Western blot analysis were used to detect the gene transcription and expression. Results PCR and Southern blot results showed that 7 of 25 mice were transgenic mice.

通过显微注射的方法,将5-脂氧化酶基因片段(6.8 kb)导入BDF1受精卵雄原核并移植到同期受孕的假孕母鼠输卵管中,对产出仔鼠的鼠尾组织DNA进行PCR、Southern blot检测,对9、20、24号转基因小鼠分别提取腹腔细胞、骨髓细胞及脾、肾组织总RNA和蛋白,并采用RT-PCR、Western blot方法进行转录水平检测和蛋白表达检测。

MTT assay FAK signaling pathway inhibitor genistein on human corneal epithelial cell cytotoxicity;RT-PCR detection of human corneal epithelial cells adhesion to fungus at different times,extracellular matrix protein including laminin,fibronectin,FN glass,Ⅳcollagen,transmembrane protein integrinαⅤ,integrinβ1(ITGβ1),as well as the FAK signaling pathway FAK1,FAK2 and Paxillin gene expression;Western blot detection of the signal transduction pathway adhesion-associated protein ITGβ1,FAK and PAX expression and the inhibition of genistein. Immunocytochemical method was used to observe the LN,FN and FAK expression in human corneal epithelial cells during interaction with the fungues;Laser scanning confocal microscope had a cell positioning on FN,FAK and PAX,observed the changing of the human corneal epithelial cytoskeleton after stimulated by fungues;Quantitatived by flow cytometry to detect of human corneal epithelial cells with PAX at ITGβ1 fungal expression after adhesion;Optical microscopy quantitied the fungues and human corneal epithelial cell adhesion and recorded to determination the integral optical density afrer adhesion;Scanning and transmitted electron microscope observed the changing of cell ultrastructure after fungues and human corneal epithelial cell adhesion.

第一部分真菌激活人角膜上皮细胞FAK信号转导通路的体外实验研究将三种常见致病真菌(白色念珠菌、烟曲霉菌和茄病镰刀菌)分别与人角膜上皮细胞共孵育,MTT法检测FAK信号通路抑制剂染料木黄酮的对人角膜上皮细胞的细胞毒性作用;RT-PCR检测真菌黏附人角膜上皮细胞后不同时间细胞外基质层连蛋白、纤连蛋白、玻连蛋白、Ⅳ型胶原、跨膜蛋白整合素αV、整合素β1(ITGβ1),以及FAK信号通路中FAK1、FAK2和桩蛋白基因的表达情况;Western blot的方法检测黏附信号转导途径相关蛋白ITGβ1、FAK和PAX的表达,以及染料木黄酮对真菌刺激人角膜上皮细胞FAK信息通路活化的抑制作用;免疫细胞化学方法观察人角膜上皮细胞与真菌相互作用过程中LN、FN和FAK的表达;激光共聚焦显微镜对FN、FAK和PAX进行了细胞定位,并观察真菌刺激后人角膜上皮细胞骨架的变化;流式细胞仪定量检测人角膜上皮细胞ITGβ1与PAX在真菌黏附后表达的改变;光学显微镜观察真菌与人角膜上皮细胞黏附数量,记录并测定了黏附后积分光密度值OD扫描及投射电镜观察了真菌与人角膜上皮细胞黏附后,细胞超微结构的改变。

The DNA of Yak PrP expressed plasmid extracted from the recombinant expressed bacterium was identified by PCR amplification and two-ezyme digestion. The expressed product was obtained from the recombinant bacterium inducted under the optimized expressed condition (37℃, 1 mmol/L IPTG, 6 h), and it was determined by SDS-PAGE and Western blotting. The GST-BoPrP(23—242) fusion proteins were collected by two ways: the first is to purify and renature from the inclusion bodies; the second is to separate by SDS-PAGE.

将保存的含有牦牛PrP基因重组表达菌提取质粒DNA,进行PCR扩增和双酶切鉴定,并在优化诱导表达条件(37℃,1mmol/L IPTG,6h)下,获得的表达产物进行SDS-PAGE和Western blotting检测;将鉴定之后的重组表达菌诱导表达后,通过两种方法回收GST-BoPrP(23~242)融合蛋白:其一,是从包涵体中提取和复性GST-BoPrP(23~242)融合蛋白;其二,是通过SDS-PAGE直接分离并纯化GST-BoPrP(23~242)融合蛋白。

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