Mohammad K, Esen A
Department of Biology, Virginia Polytechnic Institute, Blacksburg 24061.
J Immunol Methods. 1989 Feb 8;117(1):141-5. doi: 10.1016/0022-1759(89)90129-4.
The effect of blocking and non-blocking was compared in ELISA, immunostaining dot-blots and western blots using phosphate-buffered saline with and without the addition of Tween 20 for washes and dilutions. The results indicated that when Tween 20 was included in phosphate-buffered saline, a blocking agent and a blocking step were not required in immunoassays. Moreover, the use of protein-based blocking agents was found to somewhat lower the reactivity between the antibody and the antigen, possibly due to steric hindrance.
在酶联免疫吸附测定（ELISA）、免疫染色斑点印迹法和蛋白质印迹法中，使用添加和不添加吐温20的磷酸盐缓冲盐水进行洗涤和稀释，比较封闭和非封闭的效果。结果表明，当磷酸盐缓冲盐水中含有吐温20时，免疫测定中不需要封闭剂和封闭步骤。此外，发现使用基于蛋白质的封闭剂会在一定程度上降低抗体与抗原之间的反应性，这可能是由于空间位阻所致。
