| Protein-lipid Interaction Analysis by Surface Plasmon Resonance (SPR)
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Author:
Date:
2014-09-20
[Abstract] Interactions of lipids with proteins are essential events in the framework of biological membranes. Assessment of the affinity and specificity of protein-lipid binding can give useful information to elucidate cell membrane functions. Surface Plasmon Resonance (SPR) is a powerful technology to study macromolecular interactions, allowing direct and rapid determination of association and dissociation rates using small amounts of samples. An extensive range of binding analyses can be performed by SPR such as protein–protein, protein–membrane (lipids), protein–carbohydrate, protein–nucleic acid and even protein-small molecules. This protocol describes the binding of an antimicrobial protein (used as ligand) to a lipopolysaccharide (LPS) (used as analyte) after immobilization onto a CM sensor ...
[摘要] 脂质与蛋白质的相互作用是生物膜框架中的重要事件。 蛋白质 - 脂质结合的亲和力和特异性的评估可以提供有用的信息来阐明细胞膜功能。 表面等离子体共振（SPR）是研究大分子相互作用的强大技术，允许使用少量样品直接和快速测定缔合和解离速率。 可以通过SPR进行广泛范围的结合分析，例如蛋白质 - 蛋白质，蛋白质 - 膜（脂质），蛋白质 - 碳水化合物，蛋白质 - 核酸甚至蛋白质 - 小分子。 该方案描述了在通过胺偶联固定在CM传感器芯片上之后，抗微生物蛋白（用作配体）与脂多糖（LPS）（用作分析物）的结合。
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| Protein-peptide Interaction by Surface Plasmon Resonance
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Author:
Date:
2013-02-05
[Abstract] This protocol measures the protein-peptide interaction by surface plasmon resonance (SPR) using Biacore X100 (GE Healthcare). The Biacore system can monitor the direct interaction between biomolecules. There are several methods of immobilizing a ligand to the sensor chip. The optimal immobilization method for each experiment needs to be selected. In this protocol, we employed amine coupling to immobilize the protein to the carboxyl-type sensor chip. The procedure generally follows the “Instrument Handbook” of Biacore X100.
[摘要] 该协议使用Biacore X100（GE Healthcare）通过表面等离子体共振（SPR）测量蛋白质 - 肽相互作用。 Biacore系统可以监测生物分子之间的直接相互作用。 存在将配体固定到传感器芯片的几种方法。 需要选择每个实验的最佳固定方法。 在该协议中，我们使用胺偶联固定蛋白质到羧基型传感器芯片。 该程序通常遵循Biacore X100的"仪器手册"。
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