.png){kind=icon}
返回
Optimization of aptamer microarray technology for multiple protein targets
.png){kind=icon}
PRE
AI
DOI：10.1016/j.aca.2005.12.038.png)
.png){kind=icon}
原文求助.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
Aptamer-based microarrays for the quantitation of multiple protein analytes have been developed. A multiplex aptamer microarray was generated by printing two RNA aptamers (anti-lysozyme and anti-ricin) and two DNA aptamers (anti-IgE and anti-thrombin) on to either streptavidin (SA) or neutravidin (NA)-coated glass slides. However, substantial optimization was required in order to ensure the simultaneous function of the aptamer:analyte pairs. The effects of protein labeling, assay buffer. surface coating, and immobilization chemistry and orientation were investigated. A single buffer (PBS buffer containing 5 mM MgCl2 and 0.1% Tween 20) was found to work well with all the aptamers, even though this was not the buffer originally used in their selection, while neutravidin-coated slides yielded a lower detection limit, wider detection ran.-e. and more uniform background than streptavidin-coated slides. Incubation with Cy3-labeled proteins yielded sensitive, target-specific, and dose-dependent responses to each protein. Target protein concentrations as low as 72 pg/mL (5 pM, lysozyme), 15 ng/mL (0.5 nM, ricin), 1.9 ng/mL (0.01 nM, IgE), and 170 ng/mL (5 W, thrombin) Could be detected. These results show that aptarrier arrays can potentially be used with nurnerous proteins in parallel, furthering the notion that aptamer arrays may be useful in proteomics. (c) 2006 Elsevier B.V. All rights reserved.
Keyword:
aptamer
microarray
protein
multiplex
proteomics
期刊
IF:
6
论文数:
3.3W
被引数:
6.1W
机构
暂无机构信息
引用论文
分享收藏Open-tubular capillary electrochromatography of bovine β-lactoglobulin variants A and B using an aptamer stationary phase使用适体固定相对牛 β-乳球蛋白变体A和B进行开管毛细管电色谱.png)
ELECTROPHORESIS
ELECTROPHORESISIF2.5
PREAI
分享收藏Array-based multiplexed screening and quantitation of human cytokines and chemokines基于阵列的多重筛选和人类细胞因子和趋化因子的定量
PREAI
分享收藏
查看更多内容