Category:Method/Sample/GL/Acidic: Difference between revisions

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# Lipids were extracted from mouse brain with 9 ml chloroform–methanol (2:1, v/v) and then with 7.6 ml chloroform–methanol-water (1:2:0.8, v/v/v).  
# Lipids were extracted from mouse brain (500 mg) with 9 ml chloroform–methanol (2:1, v/v) and then with 7.6 ml chloroform–methanol-water (1:2:0.8, v/v/v).  
# The extraction was dried under a gentle stream of nitrogen and re-dissolved with 7.5 ml chloroform-methanol-water(30:60:8; v/v/v).
# The extraction was dried under a gentle stream of nitrogen and re-dissolved with 7.5 ml chloroform-methanol-water(30:60:8; v/v/v).
# Samples were applied to a DEAE-Sephadex A-25 column (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden).
# Samples were applied to a DEAE-Sephadex A-25 column (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden).

Revision as of 06:48, 22 October 2010

Acidic glycolipids (gangliosides, sulphatides)

  1. Lipids were extracted from mouse brain (500 mg) with 9 ml chloroform–methanol (2:1, v/v) and then with 7.6 ml chloroform–methanol-water (1:2:0.8, v/v/v).
  2. The extraction was dried under a gentle stream of nitrogen and re-dissolved with 7.5 ml chloroform-methanol-water(30:60:8; v/v/v).
  3. Samples were applied to a DEAE-Sephadex A-25 column (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden).
  4. Neutral lipids were eluted with 15 ml chloroform-methanol-water (30:60:8; v/v/v).
  5. Acidic glycolipid mixtures were eluted with 7.5 ml chloroform-methanol-0.8 M sodium acetate (30:60:8; v/v/v).

  1. マウス脳 (500 mg) からクロロホルム:メタノール混液 (2:1, v/v)9 ml, クロロホルム:メタノール:水混液 (1:2:0.8, v/v/v) 7.6 ml で抽出
  2. ゆるやかな窒素ガスで乾燥、7.5 ml クロロホルム:メタノール:水混合 (30:60:8; v/v/v) に再度溶解
  3. DEAE-Sephadex A-25 カラム (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden) にセット
  4. 中性脂質を 15 ml クロロホルム:メタノール:水混合 (30:60:8; v/v/v) で洗浄
  5. 酸性の糖脂質混合物を、 7.5 ml クロロホルム:メタノール:0.8M 酢酸ナトリウム混合 (30:60:8; v/v/v) で溶出

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