| Dictyostelium Cultivation, Transfection, Microscopy and Fractionation
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Author:
Date:
2015-06-05
[Abstract] The real time visualisation of fluorescently tagged proteins in live cells using ever more sophisticated microscopes has greatly increased our understanding of the dynamics of key proteins during fundamental physiological processes such as cell locomotion, chemotaxis, cell division and membrane trafficking. In addition the fractionation of cells and isolation of organelles or known compartments can often verify any subcellular localisation and the use of tagged proteins as bait for the immunoprecipitation of material from cell fractions can identify specific binding partners and multiprotein ...
[摘要] 使用更复杂的显微镜，活细胞中荧光标记的蛋白质的实时可视化大大增加了我们对基本生理过程如细胞运动，趋化性，细胞分裂和膜运输过程中关键蛋白质动力学的了解。此外，细胞的分级和分离细胞器或已知的隔室通常可以验证任何亚细胞定位，并且使用标记的蛋白质作为诱饵用于来自细胞部分的物质的免疫沉淀可以鉴定特异性结合配偶体和多蛋白复合物，从而有助于赋予功能标记蛋白。我们已经成功地将这些技术应用于作为古代异构六聚体膜转运复合物的一部分的盘基网柄菌discoideum蛋白TSPOON（Hirst等，2013）。 ...
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| Competitive Bone-marrow Transplantations
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Author:
Date:
2014-06-05
[Abstract] Competitive bone marrow transplantation assay measures multi-lineage reconstitution of hematopoiesis in irradiated transplant recipient mice. Thus this assay is routinely used to determine haematopoietic stem and progenitor cells (HSPCs) functionality in vivo. The principle of the method is to transplant bone marrow donor cells (derived from transgenic mice of choice) on C57BL6 background together with normal competitor bone marrow. In order to distinguish donor from competitor cells upon transplantation, usually competitor mice are congenic and carry the differential B cell antigen ...
[摘要] 竞争性骨髓移植测定测量照射的移植受体小鼠中造血的多谱系重建。 因此，该测定常规用于在体内确定造血干细胞和祖细胞（HSPC）功能。 该方法的原理是将骨髓供体细胞（源自选择的转基因小鼠）与正常竞争剂骨髓一起移植到C57BL6背景上。 为了在移植时区分供体和竞争者细胞，通常竞争性小鼠是同基因的并携带最初称为Ly5.1和CD45.1的差别B细胞抗原。典型的竞争性骨髓移植实验将含有两个移植组， 供体（选择的转基因小鼠及其对照）与正常竞争剂竞争移植，并在血液和骨髓中评价移入效率。
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