Lane J D, Zimmer H G, Neuhoff V
Hoppe Seylers Z Physiol Chem. 1979 Oct;360(10):1405-8. doi: 10.1515/bchm2.1979.360.2.1405.
A technique is presented which allows one to label and quantitate glycoproteins. Small amounts of protein from biological samples (0.5-2.5 microgram for mixtures and less for individual proteins) are separated by sodium dodecyl sulfate gel electrophoresis on 1-30% polyacrylamide gradient microgels. The gels are stained with Co-omassie Brillant Blue R250 to evaluate relative migration or fixed in 2-propanol/acetic acid and stained with fluorescein isothiocyanate-labelled concanavalin A. The microgels are then scanned using a fluorescence microscope controlled by a computer, although simpler configurations are possible. Standards of known carbohydrate composition (e.g., glucose oxidase) are used for comparative purposes. Glycoproteins in the order of 5-30 ng protein (or 1-5 ng carbohydrate) can be detected without difficulty. This technique may prove valuable in evaluating glycoproteins when the available material is limited.
本文介绍了一种能够对糖蛋白进行标记和定量的技术。生物样品中的少量蛋白质（混合物为0.5 - 2.5微克，单个蛋白质则更少）通过在1 - 30%聚丙烯酰胺梯度微型凝胶上进行十二烷基硫酸钠凝胶电泳来分离。凝胶用考马斯亮蓝R250染色以评估相对迁移情况，或者固定在异丙醇/乙酸中，并用异硫氰酸荧光素标记的伴刀豆球蛋白A进行染色。然后使用由计算机控制的荧光显微镜对微型凝胶进行扫描，不过也可以采用更简单的配置。使用已知碳水化合物组成的标准品（如葡萄糖氧化酶）用于比较目的。能够轻松检测出5 - 30纳克蛋白质（或1 - 5纳克碳水化合物）量级的糖蛋白。当可用材料有限时，这项技术在评估糖蛋白方面可能被证明是有价值的。
