Neuhoff V, Ewers E, Huether G
Hoppe Seylers Z Physiol Chem. 1981 Nov;362(11):1427-34. doi: 10.1515/bchm2.1981.362.2.1427.
A flexible method for glycoprotein determination with microliter-volumes using spot analysis ("Tüpfelprobe") on cellulose acetate layers is described. With glucose oxidase as an example of a glycoprotein and fluorescein isothiocyanate-labelled Concanavalin A, a sensitivity of 10 ng is reached; in combination with horseradish peroxidase 1 ng of glucose oxidase can be detected. A simultaneous determination of protein and glycoprotein with one single spot of 0.5, 1 or 2 microliters of a glycoprotein solution can be performed. The method is independent of many common external influences, e.g. dodecyl sulfate, Triton X-100, NP 40, mercaptoethanol and desoxycholate. Only pretreatment of the glycoprotein with urea decreases the sensitivity.
描述了一种灵活的糖蛋白测定方法，该方法使用醋酸纤维素层上的斑点分析（“点滴试验”）对微升体积的样品进行检测。以葡萄糖氧化酶作为糖蛋白的示例，并使用异硫氰酸荧光素标记的伴刀豆球蛋白A，可达到10 ng的灵敏度；与辣根过氧化物酶联合使用时，可检测到1 ng的葡萄糖氧化酶。可以用0.5、1或2微升糖蛋白溶液的单个斑点同时测定蛋白质和糖蛋白。该方法不受许多常见外部因素的影响，例如十二烷基硫酸钠、 Triton X-100、NP 40、巯基乙醇和脱氧胆酸盐。只有用尿素对糖蛋白进行预处理会降低灵敏度。
