论文部分内容阅读
以偶联磷酸化酪氨酸的牛血清白蛋白（ＢＳＡ）作为免疫原免疫兔获得抗血清．自抗血清中分离获得抗体．自酪胺合成磷酸酪胺，并偶联到溴化氰活化的Ｓｅｐｈａｒｏｓｅ４Ｂ上．抗体经磷酸酪胺－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和柱纯化，所得抗体专一性强，Ｄｏｔｂｌｏｔ显示：抗体仅对酪氨酸磷酸化的蛋白质包括酪氨酸磷酸化的血清白蛋白，溶菌酶，卵清蛋白起抗原抗体反应，而不识别非酪氨酸磷酸化的溶菌酶，卵清蛋白，也不识别作为免疫原的骨架成分ＢＳＡ，也不识别丝氨酸磷酸化的卵清蛋白和苏氨酸磷酸化的卵清蛋白．
Antiserum was obtained by immunizing rabbits with bovine serum albumin (BSA) conjugated to phosphotyrosine as an immunogen. Antibody was isolated from antiserum. Phosphotyramide was synthesized from tyramine and coupled to cyanogen bromide-activated Sepharose 4B. Antibodies were purified by phosphotyrosamine-Sepharose 4B affinity chromatography and the resulting antibodies were highly specific. Dotblot showed that the antibodies only tyrosine phosphorylated proteins include tyrosine phosphorylated serum albumin, lysozyme, ovalbumin Antigen-antibody reactivity without recognizing non-tyrosine phosphorylated lysozyme, ovalbumin, does not recognize BSA as a scaffold of the immunogen nor does it recognize serine-phosphorylated ovalbumin and threonine-phosphorylated eggs Albumin