Immunodetection of tubulin carboxypeptidase activity on nitrocellulose membrane after gel electrophoresis and blotting.

Biochemistry international Pub Date : 1992-12-01
A M Smania, C E Argaraña, J C Weizetfel, H S Barra
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Abstract

It was found that the detyrosination of tyrosinated tubulin by tubulin carboxypeptidase can occur when both the enzyme and the substrate are adsorbed on nitrocellulose. This, and the use of a specific antibody that recognizes detyrosinated tubulin allowed us to localize tubulin carboxypeptidase on a nitrocellulose membrane after agarose gel electrophoresis and blotting. The method was also extended to detect pancreatic carboxypeptidase A.

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凝胶电泳和印迹法对硝化纤维素膜微管蛋白羧肽酶活性的免疫检测。
研究发现,当微管蛋白羧肽酶和底物同时吸附在硝化纤维素上时,可以发生微管蛋白酪氨酸化的去酪氨酸化。这一点,以及识别去脱氧微管蛋白的特异性抗体的使用,使我们能够在琼脂糖凝胶电泳和印迹后将微管蛋白羧肽酶定位在硝化纤维素膜上。该方法也适用于胰羧肽酶A的检测。
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