Immunodetection of tubulin carboxypeptidase activity on nitrocellulose membrane after gel electrophoresis and blotting.
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.
Smania
A M
AM
Centro de Investigaciones en Química Biológica de Córdoba, Facultad de Ciencias Quìmicas, Universidad Nacional de Córdoba, Argentina.
Argaraña
C E
CE
Weizetfel
J C
JC
Barra
H S
HS
D000818:Animals; D001921:Brain / Q000201:enzymology; D002268:Carboxypeptidases / Q000032:analysis*; D043422:Carboxypeptidases A; D002417:Cattle; D003101:Collodion / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D015151:Immunoblotting; D007525:Isoelectric Focusing; D008567:Membranes, Artificial
D008567:Membranes, Artificial; D003101:Collodion; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; C023212:tyrosyltubulin carboxypeptidase