ELISA: suitable immunoblotting: 10-20 μg/mL using human breast cancer MCF7 cell line. immunofluorescence: 5-10 μg/mL using human foreskin Hs68 cells. immunohistochemistry: suitable
Tubulin is the major building block of microtubules. It possesses an intracellular cylindrical filamentous structure that is present in almost all eukaryotic cells. Except in the simplest eukaryotes, Tubulin exists in all cells as a heterodimer of two similar but not identical polypeptides (approx. 55 kDa each), designated α and β, that assembles into microtubules. Both α and β Tubulins undergo post-translational modifications, including acetylation, phosphorylation, detyrosination, polyglutamylation and polyglycylation. For β-tubulin, six evolutionarily conserved isotypes were identified, designated βI-βVI. Their utilization in the same cell type of different species is nearly absolutely conserved with the exception of the sequenced diversed hematopoietic β-tubulin. Since the different isotypes of tubulin differ from each other in their ability to polymerize into microtubules, it is hypnotized that the β-tubulin isotypes contribute to unique functional properties. The most complex pattern of isotype distribution in tissues is seen in the vertebrate β-tubulins. In mammals βI is constitutive and found in most tissues. βII is found in many tissues, but largely in the brain; its synthesis increases in regeneration and development of neurons. βIII is found in the brain and in dorsal root ganglia; it appears to be localized to neurons, where its expression seems to increase during axonal outgrowth. βIV in mammals exists at two subtypes, differing from each other at 10 positions. βIVa is brain specific and βIVb is ubiquitous, but both appear to be constitutive. βV is ubiquitous expressed in a variety of cultured mammalian cells. βVI is apparently restricted to hematopoietic tissues, being expressed in mammalian platelets, spleen, bone marrow and other blood-forming tissues. The detection, localization and characterization of proteins involved in microtubule function is fundamental to the understanding of mitosis, meiosis, organellar and flagellar movement, intracellular transport and cytoskeletal functions. Antibodies reacting specifically with α- and β-tubulin isotypes serve as an essential tool in the detection of the presence and functional significance of these molecules in various cellular settings.
Immunogen
Synthetic peptide corresponding to the C-terminal sequence of β-tubulin isotype IV, conjugated to BSA.
Application
Immunoblotting: a working concentration of 10-20 μg/mL is recommended using human breast cancer MCF7 cell line. Immunofluorescence: a working concentration of 5-10 μg/mL is recommended using human foreskin Hs68 cells.
Physical form
Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15mM sodium azide.
Other Notes
In order to obtain best results in different techniques and preparations we recommend determining optimal working concentration by titration test.
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