Devaraj N, Devaraj H, Bhavanandan V P
Department of Biological Chemistry and the Cell and Molecular Biology Program, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.
Anal Biochem. 1992 Oct;206(1):142-6. doi: 10.1016/s0003-2697(05)80024-7.
A procedure for the rapid isolation of mucin glycoprotein by density gradient centrifugation in cesium trifluoroacetate (CsTFA) is described. The separation of mixtures of rat tracheobronchial mucin, DNA, hyaluronic acid, and bovine serum albumin in CsTFA gradients was superior to that in cesium bromide gradients. Inclusion of guanidinium chloride or urea in the gradient had no influence on the separation obtained. The mucins isolated from sputum samples of cystic fibrosis patients by this procedure are largely free of nucleic acid, nonglycosylated proteins, and glycosaminoglycans. The results of the use of CsTFA gradient centrifugation for the isolation of mucin from extracts of bovine submaxillary gland are also presented. The CsTFA method is particularly suitable for the high-yield isolation of mucin from individual samples which are available in limited quantities.
本文描述了一种通过在三氟乙酸铯（CsTFA）中进行密度梯度离心快速分离粘蛋白糖蛋白的方法。在CsTFA梯度中分离大鼠气管支气管粘蛋白、DNA、透明质酸和牛血清白蛋白的混合物，其效果优于在溴化铯梯度中的分离效果。梯度中加入氯化胍或尿素对所得分离效果没有影响。通过该方法从囊性纤维化患者痰液样本中分离出的粘蛋白基本不含核酸、非糖基化蛋白和糖胺聚糖。文中还给出了使用CsTFA梯度离心从牛下颌下腺提取物中分离粘蛋白的结果。CsTFA方法特别适用于从少量可得的单个样本中高产率地分离粘蛋白。
