Distribution of Helium Isotopes in Western Pacific Upper Layer

3He/4He ratios in samples of sea water obtained at depths from surface to 300 m in the upper layer of the Western Pacific Ocean were measured by a mass spectrometer VG5400. The lateral and vertical distribution of He isotopes in the studied area was discussed in detail on the basis of 3He/4He ratios. Excess and depleted 3He. in relation to δ3He value of surface water in equilibrium with air has been discussed in the area, which may be mainly attributed to the incorporation of the North Pacific Intermediate Water with the Equatorial Upwelling and the exchange of water masses between the South China Sea and the Western Pacific, respectively. The present work in Western Pacific has also indicated that He isotopes may be used as a tracer for mixing processes of water masses and oceanic circulation.     

人钠/碘转运体基因转染结肠癌SW480细胞及相关蛋白表达的检测

目的以重组人钠/碘转运体（hNIS）基因转染结肠癌SW480细胞并检测hNIS mRNA及蛋白的表达，为放射性碘治疗非甲状腺肿瘤提供新思路。方法将构建好的重组质粒（pcDNA3.1+-hNIS）进行酶切、测序鉴定并扩增、提取。SW480细胞分为重组质粒（pcDNA3.1+-hNIS）转染组、空白质粒（pcDNA3.1+）转染组、空白对照组，转染后以RT-PCR和Western blot检测各组细胞hNIS mRNA和蛋白的表达。结果酶切和测序结果显示插入的hNIS基因大小和方向均正确。RT-PCR和Western blot显示重组质粒转染组SW480细胞可见hNIS mRNA和蛋白的表达，而空白对照组和空白质粒转染组均未检测到hNIS mRNA和蛋白的表达。结论脂质体法可有效地将hNIS基因转染至SW480细胞并成功表达hNIS蛋白。     

Direct Blue 71 staining as a destaining‐free alternative loading control method for Western blotting

In Western blotting, a suitable loading control is indispensable for correcting errors in the total amount of loaded protein. Immunodetection of housekeeping proteins and total protein staining have traditionally been used as loading control methods. Direct Blue 71 (DB71) staining—a novel, sensitive, dye‐binding staining method compatible with immunodetection—may offer advantages over these traditional loading control methods. Three common neuroscientific samples (human plasma, human oligodendrocytes, and rat brain) were employed to assess DB71 staining as a loading control method for Western blotting. DB71, CBB, one traditional housekeeping protein, and one protein of interest were comparatively assessed for reliability and repeatability and linear dynamic range over 2.5–40 μg of protein loaded. DB71's effect on the reliability and repeatability and linear dynamic range of immunoreaction were also assessed. Across all three sample types, DB71 was either equivalent or superior to CBB and housekeeping protein‐based methods in terms of reliability and repeatability and linear dynamic range. Across all three sample types, DB71 staining did not impair the reliability and repeatability or linear dynamic range of immunoreaction. Our results demonstrate that the DB71 staining can be used as a destaining‐free alternative loading control method for Western blotting.     

BirA*-protein A fusion protein based BioEnhancer amplifies western blot immunosignal

Western blot (protein immunoblot) is a widely used analytical technique in molecular biology. Utilizing the specific recognizing primary antibody, proteins immobilized on various matrix are investigated by subsequent visualization steps, for example, by the horse radish peroxidase conjugated secondary antibody incubation. Methods to improve the sensitivity in protein identification or quantification are appreciated by biochemists. Herein, we report a new strategy to amplify Western blot signals by constructing a probe with proximal labeling and IgG targeting abilities. The R118G mutation attenuated the biotin-AMP binding affinity of the bacterial biotin ligase BirA*, offering a proximity-dependent labeling ability, which could be used as a signal amplifier. We built a BirA*-protein A fusion protein (BioEnhancer) that specifically binds to IgG and adds biotin tags to its proximal amine groups, enhancing the immunosignal of target proteins. In our experiments, the BioEnhancer system amplified the immunosignal by tenfold compared to the standard western blot. Additionally, our strategy could couple with other signal enhancement methods to further increase the western blot sensitivity.     

A comparative study of CE-SDS,SDS-PAGE,and Simple Western: Influences of sample preparation on molecular weight determination of proteins

The development of capillary electrophoresis, especially CE-SDS devices, has led CE-SDS to become an established tool in a wide range of applications in the analysis of biopharmaceuticals and is increasingly replacing its method of origin, SDS-PAGE. The goal of this study was to evaluate the comparability of molecular weight (MW) determination especially by CE-SDS and SDS-PAGE. For ensuring comparability, model proteins that have little or no posttranslational modifications and an IgG antibody were used. Only a minor influence of sample preparation conditions, including sample buffer, temperature conditions, and different reducing agents on the MW determination were found. In contrast, the selection of the MW marker plays a decisive role in determining the accurate apparent MW of a protein. When using different MW markers, the deviation in MW determination can exceed 10%. Interestingly, CE-SDS and 10% SDS-PAGE hardly differ in their trueness of MW determination. The trueness in relation to the reference MW for each protein was calculated. Although the trueness values for the model proteins considered range between 1.00 and 1.11 using CE-SDS, they range between 0.93 and 1.03 on SDS-PAGE, depending on the experimental conditions chosen.     

A New Early Devonian Radiolarian Genus From Western Yunnan

A new radiolarian genus, Eoalbaillella belonging to Albaillellidae Deflandre emend. Holdsworth, is proposed from the Early Devonian in Western Yunnan, Southwestern China. Its test is composed of an imperforate lamellar shell and an elongate triangular framework, of which the upper part is enclosed by the lamellar shell and the lower part, without the shell, has the shape of the letter X. The relations between new genus and other radiolarian genera are studied and the type species of the genus is described.     

深海富钴结核微区X射线荧光光谱分析和数据挖掘

西太平洋富钴结核是近年来新发现的海底固体矿产资源,富含Mn,Fe,Co,Ni和Cu等多种关键金属元素。富钴结核是一种非均质的地球化学和矿物学集合体,粒径约6 cm的结核在生长过程中记录了数千万年的海洋沉积历史,亟需高分辨率的分析技术揭示古海洋环境信息。采用微区X射线荧光光谱仪（μ-XRF）,对C3BC1704富钴结核开展多元素面扫描,获得了原位高分辨率多元素的信号强度数据,评价了μ-XRF技术在富钴结核中的应用质量。元素信号谱峰特征和数据频谱分布结果显示,富钴结核中Mn,Fe,Ti,Co,Ca和Ni等元素信号强度敏感,数据呈现相对较好的正态分布特征,可用于定量或半定量分析;Si,Cu和Al等元素信号较弱,数据呈现左偏的正态分布特征,建议相关数据仅作参考。μ-XRF获得的数据量庞大且彼此独立,本研究将不同元素连接成彼此关联的多维矩阵,实现了数据的位置信息和特征元素之间的数学运算和筛选,了解了金属元素的分布和变化特征,揭示了富钴结核生长过程的环境变化。结果显示,Mn和Fe等元素在生长层中波动剧烈,金属元素在富钴结核中的分布极不均匀,显示出多成因类型的交替微层和7个大的生长周期旋回。C3BC1704富钴结核主体暴露在海水中,金属元素主要来自海水,化学组成指示为典型的水成成因。进一步定量分析结果显示,Mn,Cu和Ni等元素含量从内部向外围呈现同步降低的趋势,Fe,Ti和Co等元素含量则呈现同步升高的趋势,这些特征指示早期偏向于成岩富集环境,晚期则以水成富集为主。富钴结核金属元素的分布和变化特征,清晰呈现了富钴结核的生长结构,揭示了富钴结核生长过程的环境变化,有利于富钴结核的成矿模型的构建。     

New developments in Western blot technology

Western blotting is a highly valued method for protein identification and relative quantitation in complex samples. It combines size-based electrophoretic separation with immunoaffinity to identify specific proteins. This technique remains popular and has become a workhorse in biochemical research and clinical laboratories. Despite its utility and popularity, this method has many limitations including slow analysis, incompatibility with limited sample application, low throughput and low information content. Recently there has been significant success in improving different aspects of Western blotting. In this review, we provide an overview of the developments in the area of improving conventional Western blotting methods with a focus on recent developments in microfluidic Western blotting. We overview different separation platforms, and discuss studies on protein transfer methods as well as protein immobilization methods and chemistries. We also describe integrated miniaturized platforms that can perform rapid separations and immunodetections.