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 Protein Simple Jess 自动毛细管 电泳和蛋白质印迹系统该仪器进行蛋白质分离。 通过毛细管电泳，然后使用化学发光或凝胶内免疫印迹法 使用凝胶内荧光标记对免疫标记条带进行荧光检测。 用户还可以加载所有蛋白质的样品以及一抗和二抗。 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。 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 4 小时内，消除了蛋白质电转移固有的变异性 电泳凝胶到免疫印迹膜，灵敏度非常高（小于1.5 每个泳道需要微克蛋白质）因此，测定间变异性接近 15%，并且 因此，该技术非常适合需要如此大的实验。 需要检测的样品和/或抗体数量不断增加。 在此系统中进行了验证，目前制造商网站上列出了 1,400 多个。 仪器将促进多个领域的研究，它将用于脊髓损伤研究。 用于持续分析功能性电引起的细胞内信号传导变化 刺激正在进行的小鼠和人类受试者研究，并阐明机制 用于通过基因操作或候选药物观察到的功能结果的变化。 创伤性脑损伤研究中，这项技术对于研究不断增长的 重复后与血管和行为异常相关的蛋白质数量 在正在进行的阿尔茨海默病研究中，蛋白质水平是关键。 通过不断增长的基因组学技术确定的因素 了解载脂蛋白 epsilon 4 和载脂蛋白 epsilon 之间紧密联系的生物学基础 认知障碍的新研究将调查新候选药物。 缓解抑郁症的效果预计也将在很大程度上依赖于该系统的功能。 在未成年用户中，该仪器将用于加速多项研究的进展 细胞培养系统中癌症生物学的各个方面，持续依赖于蛋白质印迹法 各种各样的终点，并且在心血管研究中，加强以下方面的研究： 细胞内应激信号通路汇聚于缺氧诱导因子，利用 通过蛋白质印迹来表征此类途径的激活状态。