In vitro Lipid Transfer Assay
体外脂质转移试验
发布: 2013年05月05日第3卷第9期 DOI: 10.21769/BioProtoc.691 浏览次数: 12536
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参见作者原研究论文
The authors used this protocol in:
Jul 2012
Abstract
This is a protocol to detect lipid transfer activity of NRF-5, a member of the LPS binding/lipid transfer protein family. The lipid transfer activity is examined by using isotope-labeled cholesterol and liposomes, and tested in two directions (Figure 1): from proteins to liposomes and from liposomes to proteins.
Materials and Reagents
- PC (Avanti-Polar Lipids)
- PE (Avanti-Polar Lipids)
- Cholesterol (Avanti-Polar Lipids, catalog number: 700000 )
- 1,2-Dioleoyl-sn-Glycero-3-Phosphoethanolamine (DOPE) (Avanti, catalog number: 850725 )
- 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine (DOPC) (Avanti, catalog number: 770375 )
- 1 mCi (37 MBq) (PerkinElmer, catalog number: NET139001MC )
- Protein of interest tagged with Flag
- Tris-HCl (pH 7.4)
- NaCl
- Anti-Flag M2 agarose beads (Sigma-Aldrich, catalog number: A2220 )
- Flag peptide (Sigma-Aldrich, catalog number: F3290 )
- Chloroform
- Wash buffer (see Recipes)
- Elution buffer (see Recipes)
Equipment
- Avanti Mini-Extruder (Avanti, catalog number: 610023 )
- Vortexer
- Centrifuges
- Rotator
- Branson tip-sonicator (Cole-parmer Cp750)
- Scintillation counter (Wallac MicroBeta TriLux, catalog number: 1450-023 )
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Zhang, Y. and Wang, X. (2013). In vitro Lipid Transfer Assay. Bio-protocol 3(9): e691. DOI: 10.21769/BioProtoc.691.
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分类
生物化学 > 蛋白质 > 活性
生物化学 > 脂质 > 脂质转运
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