Dye Release Experiments with Dextran Loaded Vesicles
采用载有右旋糖酐的囊泡进行染色释放试验
发布: 2014年07月20日第4卷第14期 DOI: 10.21769/BioProtoc.1190 浏览次数: 10655
评审: Anonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Aug 2013
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Abstract
Dye release experiments are a widely used method to assess the interactions between membrane-active molecules and lipid membranes. Of particular interest is the ability to assess the degree of the lipid bilayer perturbation by simultaneously encapsulating dye of different sizes, such as dextrans grafted with a chromophore. In this assay, dextran linked to rhodamine or fluorescein are both encapsulated in lipid vesicles to allow quantifying the leakage of each dextran individually from a single sample. For instance, the size evaluation of the lipid pore formed by an antimicrobial peptide has been recently achieved using this protocol (Sani et al., 2013).
Keywords: Model membranes (模型膜)
Phospholipid bilayers (磷脂双分子层)
Dye leakage (染料渗漏)
Fluorescence assay (荧光测定法)
Antimicrobial peptides (抗菌肽)
Materials and Reagents
- Rhodamine-dextran (RD) of different molecular weights [for instance 40 kDa RD molecular weight (RD-40)]
- Fluorescein-dextran (FD) of different molecular weights [for instance 4.4 kDa FD (FD-4)]
- Tris.HCl
- NaCl
- MQ-water
- Triton-X100
- Peptide of interest e.g. Maculatin 1.1 (Sani et al., 2013)
- Lipids of interest e.g. 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), 1',3'-bis[1,2-dioleoyl-sn-glycero-3-phospho]-sn-glycerol (TOCL) (Sani et al., 2013)
Equipment
- Polycarbonate membrane filters (200 nm diameter)
- Extruder (for instance Avanti Mini-extruder, Alabaster)
- Bench centrifuge
- Spectrofluorimeter and quartz cuvette
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Sani, M., O’Brien-Simpson, N. M. and Separovic, F. (2014). Dye Release Experiments with Dextran Loaded Vesicles. Bio-protocol 4(14): e1190. DOI: 10.21769/BioProtoc.1190.
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分类
微生物学 > 微生物生物化学 > 脂质
生物化学 > 脂质 > 脂质结合
生物化学 > 蛋白质 > 相互作用 > 蛋白质-脂质相互作用
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