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Gradient Flotation Centrifugation of Chloroplast Membranes

叶绿体膜的梯度浮式离心实验

Venkatasalam Shanmugabalaji Venkatasalam Shanmugabalaji
FK Felix Kessler

发布: 2014年09月05日第4卷第17期 DOI: 10.21769/BioProtoc.1230 浏览次数: 11990

评审: Tie LiuAnonymous reviewer(s)

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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) search Gradient (梯度) search Chloroplast (叶绿体) search Membranes (膜) search Centrifugation (离心的) search

Materials and Reagents

  1. Anti-PGL35 (Agrisera, catalog number:  AS06 116 )
  2. Anti-TOC75 (Agrisera, catalog number:  AS06 150 )
  3. Anti-LHCB2 (Agrisera, catalog number:  AS01 003 )
  4. Na-ascorbate (Sigma-Aldrich, catalog number: 11140 )
  5. BSA fraction V (Sigma-Aldrich, catalog number: 05470 )
  6. PMSF/isopropanol (Sigma-Aldrich, catalog number: P7626 )
  7. Tricine-HCl (Sigma-Aldrich, catalog number: T0377 )
  8. DTT (Sigma-Aldrich, catalog number: 43819 )
  9. HB buffer (see Recipes)
  10. TE buffer (see Recipes)

Equipment

  1. Miracloth (pore size: 22-25 µm) (Merck KGaA, catalog number: 475855 )
  2. Centrifuge with JA-14 rotor
  3. Potter homogenizer
  4. Polycarbonate UltraClear SW28 tube
  5. SW41Ti rotor (Beckman Coulter)
  6. Spectrophotometer

Procedure

English
中文翻译

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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