An assay method for ganglioside synthase using anion-exchange chromatography.

An assay method for ganglioside synthase using anion-exchange chromatography.
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使用阴离子交换色谱法测定神经节苷脂合酶的方法。

DOI:
10.1016/0003-2697(89)90709-4
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发表时间:
1989
影响因子:
2.9
通讯作者:
A. Makita
A. Makita
中科院分区:
生物学4区
文献类型:
--
作者:
M. Kawano;K. Honke;M. Tachi;S. Gasa;A. Makita

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建立了一种基于离子交换色谱和溶解度相结合的快速测定神经节苷脂酶和神经节苷脂磺基转移酶活性的方法。该方法包括在DEAE-Sephadex柱上选择性洗脱不含标记前体和分解产物的放射性标记反应产物(酸性糖脂),以及对洗脱液中的产物进行直接放射性测定。用40 mm乙酸铵(AcONH 4)的甲醇溶液从柱上洗脱单唾液酸神经节苷脂,用90 mm AcONH 4的甲醇溶液从柱上洗脱硫酸唾液酸神经节苷脂,用40 mm AcONH 4的异丙醇/正己烷/水(55/20/19,v/v/v)从柱上洗脱双唾液酸神经节苷脂。所建立的方法简单、重复性好、经济。以大鼠高尔基体膜为酶源,产品回收率达95%以上。
A rapid procedure which is based on combined ion-exchange chromatography and solubility was established for determination of the activity of ganglioside synthases and cerebroside sulfotransferase. The procedure consists of selective elution of radiolabeled reaction products (acidic glycolipids) freed from labeled precursors and breakdown products on a DEAE-Sephadex column and of direct radioassay of the products in the eluate. Monosialogangliosides were eluted from the column with 40 mm ammonium acetate (AcONH4) in methanol, cerebroside sulfate with 90 mm AcONH4in methanol, and disialogangliosides with 40 mm AcONH4in isopropanol/n-hexane/water (55/20/19, v/v/v). The established procedure is simple, reproducible, and economical. Using rat Golgi membrane as enzyme source the recovery rate of the products was over 95%.
DOI: --
发表时间: 1985
影响因子: 6.5
作者:
D. Figlewicz;C. Nolan;I. Singh;F. B. Jungalwala
通讯作者: D. Figlewicz;C. Nolan;I. Singh;F. B. Jungalwala
使用离子交换色谱法进行神经节苷脂糖基转移酶测定。
DOI: 10.1016/0003-2697(86)90134-x
发表时间: 1986
影响因子: 2.9
作者:
Walton,KM;Schnaar,RL
通讯作者: Schnaar,RL
DOI: --
发表时间: 1981
期刊: The Journal of biological chemistry
影响因子: --
作者:
Reitman,ML;Kornfeld,S
通讯作者: Kornfeld,S