Gradient Flotation Centrifugation of Chloroplast Membranes
叶绿体膜的梯度浮式离心实验
发布: 2014年09月05日第4卷第17期 DOI: 10.21769/BioProtoc.1230 浏览次数: 11232
评审: Tie LiuAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Jan 2013
Abstract
Plastoglobules are lipoprotein particles physically attached to thylakoids in chloroplasts (Kessler et al., 1999). They are mainly composed of polar lipid, neutral lipids, and proteins (Vidi et al., 2006). Here we used simple sucrose gradient flotation centrifugation method to purify the plastoglobules from total chloroplast membranes (Vidi et al., 2007, Shanmugabalaji et al., 2013).
Keywords: Plastoglobule (plastoglobule)
Gradient (梯度)
Chloroplast (叶绿体)
Membranes (膜)
Centrifugation (离心的)
Materials and Reagents
- Anti-PGL35 (Agrisera, catalog number: AS06 116 )
- Anti-TOC75 (Agrisera, catalog number: AS06 150 )
- Anti-LHCB2 (Agrisera, catalog number: AS01 003 )
- Na-ascorbate (Sigma-Aldrich, catalog number: 11140 )
- BSA fraction V (Sigma-Aldrich, catalog number: 05470 )
- PMSF/isopropanol (Sigma-Aldrich, catalog number: P7626 )
- Tricine-HCl (Sigma-Aldrich, catalog number: T0377 )
- DTT (Sigma-Aldrich, catalog number: 43819 )
- HB buffer (see Recipes)
- TE buffer (see Recipes)
Equipment
- Miracloth (pore size: 22-25 µm) (Merck KGaA, catalog number: 475855 )
- Centrifuge with JA-14 rotor
- Potter homogenizer
- Polycarbonate UltraClear SW28 tube
- SW41Ti rotor (Beckman Coulter)
- Spectrophotometer
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Shanmugabalaji, V. and Kessler, F. (2014). Gradient Flotation Centrifugation of Chloroplast Membranes. Bio-protocol 4(17): e1230. DOI: 10.21769/BioProtoc.1230.
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分类
植物科学 > 植物细胞生物学 > 细胞器分离
植物科学 > 植物生理学 > 光合作用
细胞生物学 > 细胞器分离 > 叶绿体
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