ShEEP-IC: Jess Simple Western System

SheEEP-IC：Jess 简单西方系统

• 批准号: 10176059
• 负责人: CHRISTOPHER P CARDOZO
• 金额: --
• 依托单位: JAMES J PETERS VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176059
• 关键词: Alzheimer' s Disease; Antibodies; Apolipoproteins; Area; Behavioral; Biological; Biological Assay; Biology of Aging; Blood Vessels; Blood capillaries; Cancer Biology; Capillary Electrophoresis; Cardiovascular system; Cell Culture System; Cells; Computers; Data; Detection; Exposure to; Fluorescence; Funding; Gel; Genomics; Health; Hour; Hypoxia Inducible Factor; Image; Immunoblotting; Impaired cognition; Knowledge; Label; Laboratories; Link; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Manufacturer Name; Medical; Membrane; Mental Depression; Mental Health; Methods; Minor; Modeling; Mus; Pathway interactions; Productivity; Proteins; Quality of life; Reproducibility; Request for Applications; Research; Research Personnel; Running; Sampling; Sheep; Signal Pathway; Signal Transduction; Site; Speed; Spinal cord injury; Stress; System; Techniques; Technology; Tissues; Training; Traumatic Brain Injury; Tube; Veterans; Western Blotting; drug candidate; experimental study; functional electrical stimulation; functional outcomes; gel electrophoresis; genetic manipulation; human subject; improved; instrument; link protein; new technology; novel therapeutics; web site

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.

该申请要求资金购买蛋白质简单的Jess自动毛细管 电泳和蛋白质印迹系统。该仪器执行蛋白质分离 通过毛细管电泳，然后用化学发光或 免疫标记带的荧光检测。使用凝胶荧光标记进行标准化 在所有蛋白质中，也可用。用户加载样品以及初级和次要抗体 进入多壁板。从蛋白质加载到蛋白质分离的所有其他步骤， 免疫检测和图像采集是自动完成的。特定优势 技术包括获得24个样本的蛋白质印迹和标准化数据的能力 在不到4个小时的时间内消除了蛋白质电转移固有的变异性 电泳凝胶可免疫印迹膜和非常高灵敏度（小于1.5 微克蛋白是每条车道需要的）。结果，测定间的可变性接近15％，并且 吞吐量增加。因此，该技术非常适合实验，要求大型 样品和/或抗体的数量运行。越来越多的抗体已经 在此系统中进行了验证，目前在制造商网站上列出了1,400多个。这 仪器将促进多个领域的研究。在脊髓损伤研究中，将使用 持续分析功能电子引起的细胞内信号传导的变化 刺激小鼠和人类受试者正在进行的研究，并阐明机制 对于使用遗传操纵或候选药物观察到的功能结果的变化。 创伤性脑损伤研究，这项技术对于研究增长将是无价的 重复后，与血管和行为异常相关的蛋白质数量 暴露于轻微的爆炸波。在正在进行的阿尔茨海默氏病研究中，关键的蛋白质水平 通过不断增长的基因组技术范围识别的因素 了解载脂蛋白Epsilon 4和 认知障碍将受到质疑。调查新毒品的新研究 预计抑郁症的影响也很大程度上依赖于该系统的能力。 在次要用户中，该仪器将用于加速多重研究的进度 癌症生物学在细胞培养系统中的各个方面，不断依赖于蛋白质印迹 在心血管研究中，各种各样的终点，以增强重点的研究 在利用缺氧诱导因素上收敛的细胞内应力信号通路上 Western印迹以通过蛋白质印迹来表征这种途径的激活状态。