首页> 外文会议>International Conference on Miniaturized Systems for Chemistry and Life Sciences >ON-CHIP WESTERN BLOTTING: IN-SITU RENATURATION OF SDS-PROTEIN COMPLEXES UNIFIES SODIUM DODECYL SULFATE (SDS) SIZING BLOTTING IN ONE MICRODEVICE
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ON-CHIP WESTERN BLOTTING: IN-SITU RENATURATION OF SDS-PROTEIN COMPLEXES UNIFIES SODIUM DODECYL SULFATE (SDS) SIZING BLOTTING IN ONE MICRODEVICE

机译:片上蛋白质印迹:原位复合SDS-蛋白质复合物统一在一个微型内部的十二烷基硫酸钠(SDS)尺寸和印迹

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Western blotting readily identifies specific proteins amidst complex biological backgrounds[l, 2]. Nevertheless, immunoblotting suffers from tremendous labor-intensive and time-intensive requirements [3]. The slab-gel assays require 1-2 days for completion with multiple hands-on "blotting" steps and yield semi-quantitative information. Recently, our group has introduced new approaches for completing Western blotting. The microfluidic integration strategies introduced and used allow rapid results reporting, full assay automation, and limited sample consumption (1-10 uL). Our integration strategies use spatial, temporal, and spatiotemporal modulation of separation mechanisms in fully electrophoretic systems. The present study reports on recapitulation of immunoaffinity in previously sized proteins, using novel in-transit electrophoretic removal of SDS from SDS-protein complexes. Early results show both the length- and timescales for protein 'renaturation' are compatible with on-chip operation. Further, substantial binding affinity is recapitulated using this streamlined and promising approach.
机译:Western Blotting容易识别复杂生物背景中的特异性蛋白质[L,2]。尽管如此,免疫印迹遭受了巨大的劳动密集型和时间 - 密集的要求[3]。平板凝胶测定需要1-2天,以便完成多动属“印迹”步骤并收益半量值信息。最近,我们的小组推出了完成Western Blotting的新方法。介绍和使用的微流体集成策略允许快速结果报告,全部测定自动化和有限的样品消耗(1-10μl)。我们的集成策略在完全电泳系统中使用空间,时间和时空调制分离机制。本研究报告了在先前大小的蛋白质中,使用来自SDS-蛋白复合物的新型电泳除去SDS的In-Transit电泳除去了预先进行了免疫亲和力。早期结果表明,蛋白质“复归”的长度和时间尺度都与片上操作相容。此外,使用这种流线型和有前途的方法重新携带具有实质结合亲和力。

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