Developing a sensitive and quantitative western blot kit

开发灵敏的定量蛋白质印迹试剂盒

• 批准号: 10256088
• 负责人: Hong Yan Liu
• 金额: $ 25.6万
• 依托单位: DOTQUANT, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10256088
• 关键词: Address; Affect; Animals; Antibodies; Attenuated; Biochemical Reaction; Biological; Biological Assay; Biology; Cells; Color; Communities; Complement; Data; Detection; Disadvantaged; Dyes; ERBB2 gene; Elastomers; Enzyme-Linked Immunosorbent Assay; Enzymes; Evaluation; Feedback; Fluorescence; Future; Gel; Goals; Gold; Horseradish Peroxidase; Hybrids; Image; Immunoblotting; Investigation; Kinetics; Label; Life; Light; Medicine; Membrane; Methods; Microscopy; Modeling; Modernization; Optics; Outcome; Phase; Photobleaching; Photons; Protein Analysis; Proteins; Protocols documentation; Quantum Dots; Reaction; Reproducibility; Research; Sampling; Signal Transduction; Source; Spottings; Standardization; Streptavidin; Techniques; Technology; Testing; Time; Tissues; Transillumination; Visualization; Western Blotting; absorption; base; design; detection method; detection sensitivity; flexibility; fluorescence microscope; fluorophore; imager; imaging probe; imaging system; improved; instrumentation; interest; member; migration; next generation; novel; particle; phase 2 study; polyvinylidene fluoride; technology development

ABSTRACT Western blotting is one of the most widely used bioanalytical techniques for the detection of proteins of interest in an unknown mixture such as cell/tissue homogenate or extract. This technology was invented in the late 1970s and has enabled numerous discoveries in biology and medicine in the past 40 years. Although it is not in the spotlight anymore, western blotting has been applied at least two times more often than other competing or complementing techniques, such as mass spec, ELISA, and IHC within the last 10 years. Despite this amazingly wide acceptance by the scientific community, western blotting, in general, is not a very sensitive technique and is only semi-quantitative. The lack of quantitation in Western blot has raised major concerns in the scientific community on data reproducibility. Building on two breakthroughs we made in recent years, monovalent Qdot probes and optically clear transfer membranes, we propose to develop the next generation Western blot kits with improved sensitivity, quantification ability, multiplexing capability and flexibility in readout (standard gel UV imager or microscopy-based particle counting). If successful, we expect this technology to quickly become a commercial product to power biology research for another 40 years. .

抽象的 蛋白质印迹是用于检测感兴趣蛋白质的最广泛使用的生物分析技术之一 在未知的混合物中，例如细胞/组织匀浆或提取物。这项技术是在最近发明的 1970年代，在过去的40年中，在生物学和医学方面有许多发现。虽然不是 在聚光灯下，蛋白质印迹的应用比其他竞争的频率高两倍 或补充技术，例如群众规格，ELISA和IHC，在过去的10年内。尽管如此 一般而言，科学界的蛋白质印迹广泛接受并不是一个非常敏感的 技术，只是半定量的。缺乏蛋白质印迹的定量引起了主要的关注 有关数据可重复性的科学界。以我们近年来取得的两个突破为基础 单价QDOT探针和光学清晰的转移膜，我们建议开发下一代 蛋白质印迹套件具有提高灵敏度，定量能力，多路复用能力和读数的灵活性 （标准凝胶紫外成像仪或基于显微镜的粒子计数）。如果成功，我们希望这项技术能够 迅速成为启动生物学研究的商业产品，已有40年了。 。