Hoffman W L, Jump A A
J Immunol Methods. 1986 Nov 20;94(1-2):191-6. doi: 10.1016/0022-1759(86)90232-2.
It has generally been assumed that the binding of most proteins to nitrocellulose is stable in the non-ionic detergent Tween 20. However, the following paper demonstrates that in immunoassays where antibodies or other proteins are bound directly to the nitrocellulose, 0.05% Tween 20 may dissociate these proteins from the membrane. The degree of dissociation appears to be dependent on the individual protein studied. Some antibodies and other proteins bind tightly to nitrocellulose and dissociation of these proteins by Tween 20 is barely detectable. In contrast, other proteins are nearly completely stripped from the nitrocellulose by the same detergent. Therefore, unless it is known from control experiments what proteins will or will not be dissociated from nitrocellulose by Tween 20, the routine use of Tween 20 in the development of Western blots, native blots and dot-blots should be discontinued.
人们通常认为，大多数蛋白质与硝酸纤维素的结合在非离子去污剂吐温20中是稳定的。然而，以下论文表明，在抗体或其他蛋白质直接结合到硝酸纤维素的免疫测定中，0.05%的吐温20可能会使这些蛋白质从膜上解离。解离程度似乎取决于所研究的单个蛋白质。一些抗体和其他蛋白质与硝酸纤维素紧密结合，吐温20对这些蛋白质的解离几乎无法检测到。相比之下，其他蛋白质几乎被相同的去污剂从硝酸纤维素上完全剥离。因此，除非通过对照实验知道哪些蛋白质会或不会被吐温20从硝酸纤维素上解离，否则应停止在蛋白质免疫印迹、天然印迹和斑点印迹检测中常规使用吐温20。
