ShEEP-IC: Jess Simple Western System

此申请要求资金购买蛋白质简单杰斯自动毛细管 电泳和蛋白质印迹系统。这台仪器可以分离蛋白质 通过毛细管电泳，然后用荧光或免疫印迹法进行凝胶内免疫印迹， 免疫标记条带的荧光检测。使用凝胶内荧光标记进行归一化 所有的蛋白质都是可用的。用户加载样本以及一抗和二抗 放入多孔板中。从蛋白质装载到蛋白质分离的所有其他步骤， 免疫检测和图像采集自动完成。具体优势 该技术包括获得24个样品的蛋白质印迹和标准化数据的能力 在4小时内，消除了蛋白质从电转移中固有的可变性， 电泳凝胶免疫印迹膜，和非常高的灵敏度（小于1.5 每泳道需要微克蛋白质）。因此，试验间变异性接近15%， 增加了吞吐量。因此，这项技术非常适合需要如此大的 运行大量的样品和/或抗体。越来越多的抗体被 在这个系统中得到验证，目前在制造商的网站上列出了1,400多个。的 仪器将促进若干领域的研究。在脊髓损伤研究中， 用于持续分析由功能性电刺激引起的细胞内信号传导的变化， 在小鼠和人类受试者正在进行的研究中的刺激，并阐明机制 基因操作或候选药物观察到的功能结果的变化。在 创伤性脑损伤的研究，这项技术将是非常宝贵的，在调查日益增长的 许多蛋白质与血管和行为异常有关， 暴露在轻微的冲击波中在正在进行的阿尔茨海默病研究中，蛋白质水平的关键 通过不断增长的基因组学技术确定的因素， 了解载脂蛋白<$4与 将询问认知障碍。新的研究调查新的候选药物， 减轻抑郁症的影响预计也将在很大程度上依赖于这一系统的能力。 在少数用户中，该工具将用于加快对多种生物多样性的研究进展。 细胞培养系统中的癌症生物学方面持续依赖于蛋白质印迹， 各种各样的终点，并在心血管研究，以加强研究，重点 集中于缺氧诱导因子的细胞内应激信号通路， 蛋白质印迹法以通过蛋白质印迹法表征这些途径的活化状态。

This application requests funds to purchase a Protein Simple Jess automated capillary electrophoresis and western blotting system. This instrument performs separation of proteins by capillary electrophoresis followed by in-gel immunoblotting with chemiluminescent or fluorescent detection of immunolabeled bands. Normalization using in-gel fluorescent labeling of all proteins is also available. The user loads samples and primary and secondary antibodies into a multiwell plate. All other steps from protein loading to protein separation, immunodetection and image acquisition are done automatically. Specific advantages of this technology include the ability to obtain western blotting and normalization data for 24 samples in under 4 hours, elimination of the variability inherent in electro-transfer of protein from the electrophoresis gel to immunoblotting membranes, and very high sensitivity (less than 1.5 micrograms protein is required per lane). As a result, inter-assay variability approaches 15% and throughput is increased. This technology is thus well suited to experiments requiring that large numbers of samples and/or antibodies be run. A growing number of antibodies have been validated in this system, with over 1,400 currently listed on the manufacturer’s website. The instrument will facilitate research in several areas. In spinal cord injury research, it will be used for ongoing analysis of changes in intracellular signaling elicited by functional electrical stimulation in ongoing research in both mice and human subjects, and to elucidate mechanisms for changes in functional outcomes observed with genetic manipulation or candidate drugs. In traumatic brain injury research, this technology will be invaluable in investigating a growing number of proteins linked to vascular and behavioral abnormalities following repeated exposures to mild blast waves. In ongoing Alzheimer’s disease research, protein levels of key factors identified through an ever-growing range of genomics techniques aimed at understanding the biological basis for the strong link between apolipoprotein epsilon 4 and cognitive impairment will be interrogated. New research investigating new drug candidates to mitigate effects of depression is also expected to rely heavily on the capabilities of this systems. Among minor users, the instrument will be used to accelerate progress on research on multiple aspects of cancer biology in cell culture systems that relies continuously on western blotting for a wide variety of endpoints and, in cardiovascular research, to enhance research that focuses on intracellular stress signaling pathways converging on hypoxia-inducible factors which utilizes western blotting to characterize activation states of such pathways by western blotting.