Category:Method/Sample/GL/Acidic: Difference between revisions
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=Acidic glycolipids (gangliosides, sulphatides)= | =Acidic glycolipids (gangliosides, sulphatides)= | ||
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# Lipids were extracted from mouse brain with chloroform–methanol (2:1, v/v) and then with 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). | ||
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# 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). | ||
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# | # 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 | ||
# | # 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) | # 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 | # 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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Latest revision as of 08:17, 5 November 2010
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Acidic glycolipids (gangliosides, sulphatides)
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