SILVER STAINING OF EXTENSIVELY GLYCOSYLATED PROTEINS ON SODIUM DODECYL-SULFATE POLYACRYLAMIDE GELS - ENHANCEMENT BY CARBOHYDRATE-BINDING DYES
SILVER STAINING OF EXTENSIVELY GLYCOSYLATED PROTEINS ON SODIUM DODECYL-SULFATE POLYACRYLAMIDE GELS - ENHANCEMENT BY CARBOHYDRATE-BINDING DYES
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DOI：
10.1016/0003-2697(90)90302-p
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发表时间：
1990-03-01
影响因子：
2.9
通讯作者：
JAHNKE, MR
中科院分区：
文献类型：
--
作者：
JAY, GD;CULP, DJ;JAHNKE, MR
描述了两种用于在十二烷基硫酸钠-聚丙烯酰胺凝胶上检测小于1μg的高度糖基化蛋白质（例如粘蛋白）的方法。它们将常用的高碘酸希夫 (PAS) 和阿尔新蓝染料与银染色相结合。碳水化合物预染色使粘蛋白更具阳离子性，并有利于与离子银更好的络合。对用 PAS-银或阿尔新蓝-银染色的不同粘蛋白样品进行比较表明两种技术之间的染色差异。这种差异可能部分归因于阿尔新蓝对硫酸化糖蛋白的亲和力。这两种染色方案单独与银染色结合使用时，对于评估样品纯度和在粘蛋白纯化方案期间检测污染蛋白质特别有价值。
Two methods are described for detecting less than 1 .mu.g of highly glycosylated proteins, such as mucins, on sodium dodecyl sulfate-polyacrylamide gels. They combine commonly employed periodic acid-Schiff (PAS) and Alcian blue dyes with silver stain. Carbohydrate prestaining renders mucins more cationic and favors greater complexation with ionic silver. Comparisons of different mucin samples stained either with PAS-silver or alcian blue-silver indicate differential staining between the two techniques. Such differences may, in part, be due to an affinity of Alcian blue for sulfated glycoproteins. These two staining protocols when used in conjunction with silver staining alone are particularly valuable for assessing sample purity and for detecting contaminating proteins during mucin purification protocols.