EFFECTS OF THE BLOCKING-AGENTS BOVINE SERUM-ALBUMIN AND TWEEN 20 IN DIFFERENT BUFFERS ON IMMUNOBLOTTING OF BRAIN PROTEINS AND MARKER PROTEINS
EFFECTS OF THE BLOCKING-AGENTS BOVINE SERUM-ALBUMIN AND TWEEN 20 IN DIFFERENT BUFFERS ON IMMUNOBLOTTING OF BRAIN PROTEINS AND MARKER PROTEINS
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DOI：
10.1016/0022-1759(86)90010-4
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发表时间：
1986-01-01
影响因子：
2.2
通讯作者：
SVENNEBY, G
中科院分区：
文献类型：
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作者：
WEDEGE, E;SVENNEBY, G
比较了两种不同抗血清在pH值为7.2和10.2的缓冲液中阻断剂牛血清白蛋白和Tween 20的免疫印迹作用。抗血清分别针对来自人脑、可溶于高氯酸的纯化脑特异性蛋白和来自猪脑的磷酸盐激活谷氨酰胺酶。抗原为粗高氯酸可溶性脑提取物、粗脑磷酸盐激活谷氨酰胺酶组分和常用的分子量标记蛋白。两种抗血清与相应脑抗原制剂的结合模式不同，取决于阻断剂和阻断缓冲液的pH。此外，一些阻断缓冲液还观察到抗体与分子量标记蛋白的结合。用tris -生理盐水（pH 10.2，含有3%牛血清白蛋白作为阻断剂和抗血清稀释缓冲液）进行免疫印迹，发现抗体与标记蛋白的结合可忽略，与脑抗原的结合最特异性。
The effects of the blocking agents bovine serum albumin and Tween 20 in buffers at pH values 7.2 and 10.2 were compared in immunoblotting with 2 different antisera. The antisera were raised against a purified brain-specific protein fraction from human brain, soluble in perchloric acid, and phosphate-activated glutaminase from pig brain, respectively. The antigens were a crude perchloric acid-soluble brain extract, a crude brain phosphate-activated glutaminase fraction, and proteins commonly used as molecular weight markers. The binding patterns of the 2 antisera to the respective brain antigen preparations changed, depending on the blocking agent and the pH of the blocking buffer. Also, antibody binding to the molecular weight marker proteins was observed with some of the blocking buffers. Immunoblotting with Tris-saline, pH 10.2 containing 3% bovine serum albumin as blocking agent and diluting buffer for the antisera, showed negligible antibody binding to the marker proteins and most specific binding to the brain antigens.