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

(New page: =Acidic glycolipids (gangliosides, sulphatides)= {{Twocolumn| # Lipids were extracted with chloroform–methanol (2:1, v/v) and chloroform–methanol-water (1:2:0.8, v/v/v). # The extrac...)
 
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=Acidic glycolipids (gangliosides, sulphatides)=
=Acidic glycolipids (gangliosides, sulphatides)=


{{Twocolumn|
{{Twocolumn|
# Lipids were extracted with chloroform–methanol (2:1, v/v) and 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).
# Wash neutral lipids from the column with 15 ml chloroform-methanol-water (30:60:8; v/v/v)
# Neutral lipids were eluted with 15 ml chloroform-methanol-water (30:60:8; v/v/v).
# Acidic glycolipid mixtures were eluted with 7.5 ml chloroform-methanol-0.8 M sodium acetate (30:60:8; v/v/v).
# Acidic glycolipid mixtures were eluted with 7.5 ml chloroform-methanol-0.8 M sodium acetate (30:60:8; v/v/v).
|
|
# クロロホルム:メタノール混合 (2:1, v/v), クロロホルム:メタノール:水混合 (1:2:0.8, v/v/v) で抽出
# 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 で脂質を抽出
# 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
# 窒素ガス下で乾固、クロロホルム:メタノール:水混液 (30:60:8; v/v/v) 7.5 ml に再度溶解
# DEAE-Sephadex A-25 カラム (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden) にセット
# DEAE-Sephadex A-25 カラム (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden) に注入
# 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
# 中性脂質を クロロホルム:メタノール:水混合 (30:60:8; v/v/v) 15 ml で溶出
# 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
# 酸性糖脂質を、 クロロホルム:メタノール:0.8M 酢酸ナトリウム混液 (30:60:8; v/v/v) 7.5 ml で溶出
}}
}}

Latest revision as of 08:17, 5 November 2010

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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 で脂質を抽出
  2. 窒素ガス下で乾固、クロロホルム:メタノール:水混液 (30:60:8; v/v/v) 7.5 ml に再度溶解
  3. DEAE-Sephadex A-25 カラム (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden) に注入
  4. 中性脂質を クロロホルム:メタノール:水混合 (30:60:8; v/v/v) 15 ml で溶出
  5. 酸性糖脂質を、 クロロホルム:メタノール:0.8M 酢酸ナトリウム混液 (30:60:8; v/v/v) 7.5 ml で溶出

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