Kondo M, Harada H, Sunada S, Yamaguchi T
Honen Corporation Research Laboratories, Shimizu, Japan.
Electrophoresis. 1991 Sep;12(9):685-6. doi: 10.1002/elps.1150120918.
We increased selectivity in the detection of glycoproteins on nitrocellulose membranes by introducing a washing step using sodium hydroxide solution. Glycoproteins on the nitrocellulose membrane were first oxidized by sodium periodate; biotin hydrazide was then coupled to the aldehyde groups generated in the sugar moiety of the glycoproteins. The membrane was washed twice using sodium hydroxide solution, and avidin-horseradish peroxidase was then coupled to the remaining biotin. This system allows the detection of nanograms of glycoproteins on nitrocellulose membranes, and its specificity allows the clear distinction of glycoproteins from the nonglycosylated protein of bovine serum albumin.
我们通过引入使用氢氧化钠溶液的洗涤步骤，提高了在硝酸纤维素膜上检测糖蛋白的选择性。硝酸纤维素膜上的糖蛋白首先用过碘酸钠氧化；然后将生物素酰肼与糖蛋白糖部分产生的醛基偶联。用氢氧化钠溶液将膜洗涤两次，然后将抗生物素蛋白-辣根过氧化物酶与剩余的生物素偶联。该系统能够检测硝酸纤维素膜上纳克级的糖蛋白，其特异性使得能够将糖蛋白与牛血清白蛋白的非糖基化蛋白清晰区分开来。
