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CST1 antibody specifically recognizes cold stable tubulin
Author(s) -
Wray R.,
Szebenyi G.,
Brady S. T.
Publication year - 2002
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.81.s1.21_2.x
Subject(s) - immunostaining , biology , tubulin , western blot , blot , antibody , monoclonal antibody , axon , microbiology and biotechnology , population , clone (java method) , immunohistochemistry , microtubule , biochemistry , immunology , dna , medicine , environmental health , gene
Cold stable tubulin (CST) is isolated by a cold calcium fractionation method that results in a cold soluble fraction and a stable cold insoluble fraction. Previous biochemical studies suggest that a post‐translational modification, polyamination, is responsible for the stable form of microtubules. CST is enriched in the adult central nervous system. However, because of a mixed population of cells in biochemical studies, little is known about CST's cellular distribution. Without an antibody against CST, the cellular localization and tissue distribution could not be determined definitively. Therefore, we developed a monoclonal antibody to CST. The antibody was raised and screened against the gel purified cold insoluble fraction. Its specificity for CST was then tested using ELISA, Western blotting and immunostaining. One clone (CST1) specifically recognized the cold insoluble but not the cold soluble microtubule fraction on both Western blot and ELISA. With immunostaining in the adult rat brain, reactivity was seen in both cortical neurons and axon tracks. The staining was restricted to a subset of NF‐L positive neurons. The signal in projection neurons was enriched in cell bodies and proximal axons, but was not detected in MAP2 positive dendrites. With this novel probe we can now gain a better understanding of CST's distribution and function in the CNS.