.png){kind=icon}
返回
Microfluidic Western blotting
.png){kind=icon}
OA
AI
DOI：10.1073/pnas.1207754110.png)
.png){kind=icon}
.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
Rapid, quantitative Western blotting is a long-sought bioanalytical goal in the life sciences. To this end, we describe a Western blotting assay conducted in a single glass microchannel under purely electronic control. The mu Western blot is comprised of multiple steps: sample enrichment, protein sizing, protein immobilization (blotting), and in situ antibody probing. To validate the microfluidic assay, we apply the mu Western blot to analyses of human sera (HIV immunoreactivity) and cell lysate (NF kappa B). Analytical performance advances are achieved, including: short durations of 10-60 min, multiplexed analyte detection, mass sensitivity at the femtogram level, high-sensitivity 50-pM detection limits, and quantitation capability over a 3.6-log dynamic range. Performance gains are attributed to favorable transport and reaction conditions on the microscale. The multistep assay design relies on a photopatternable (blue light) and photoreactive (UV light) polyacrylamide gel. This hydrophilic polymer constitutes both a separationmatrix for protein sizing and, after brief UV exposure, a protein immobilization scaffold for subsequent antibody probing of immobilized protein bands. We observe protein capture efficiencies exceeding 75% under sizing conditions. This compact microfluidic design supports demonstration of a 48-plex mu Western blot in a standard microscope slide form factor. Taken together, the mu Western blot establishes a foundation for rapid, targeted proteomics by merging exceptional specificity with the throughput advantages of multiplexing, as is relevant to a broad range of biological inquiry.
Keyword:
immunoblotting
medical diagnostics
protein microarrays
systems biology
electrophoresis
AI总结
对已上传原文的论文进行重点信息的提取，主要内容包括：简要概述、研究摘要、背景介绍、关键亮点、图文解析、展望与总结。
期刊
P
IF:
9.1
论文数:
10.8W
被引数:
73.5W
机构
引用论文
Long-term control of Chagas disease in Venezuela: effects on serologic findings, electrocardiographic abnormalities, and clinical outcome..png)
Circulation
CirculationIF0
PREAI
分享收藏Homogeneous Immunosubtraction Integrated with Sample Preparation Enabled by a Microfluidic FormatANALYTICAL CHEMISTRYOAAI
ANALYTICAL CHEMISTRYIF6.7
OAAI分享收藏
查看更多内容