| Clone: | TU-08 |
|---|---|
| Isotype: | Mouse IgM |
| Specificity: | The antibody TU-08 recognizes alpha, beta-tubulin heterodimer (porcine brain), a basic intracellular structural unit of microtubules. Alpha- and beta- tubulins form approximately 100 kDa tubulin heterodimer, a globular protein that polymerizes to form microtubules. |
| Immunogen: | Microtubule proteins from porcine brain. |
| Species Reactivity: | Human, Porcine, Mouse, Other not tested |
| Application: |
Western Blotting
Recommended dilution: 5 μg/ml
Positive control: porcine brain lysate Sample preparation: Mix lysate with reducing Laemmli SDS-PAGE sample buffer. Application note: Reducing conditions.
Immunocytochemistry
|
| Purity: | > 95 % |
| Purification: | Purified from ascites by precipitation methods and ion exchange chromatography. |
| Concentration: | 1 mg/ml |
| Storage Buffer: | Tris buffered saline (TBS) with 15 mM sodium azide, approx. pH 8.0 |
| Storage / Stability: | Store at 2-8°C. Do not use after expiration date stamped on vial label. Do not freeze. |
| Expiration: | See vial label |
| Lot Number: | See vial label |
| Background: | The microtubules are intracellular dynamic polymers made up of evolutionarily conserved polymorphic alpha/beta-tubulin heterodimers and a large number of microtubule-associated proteins (MAPs). The microtubules consist of 13 protofilaments and have an outer diameter 25 nm. Microtubules have their intrinsic polarity; highly dynamic plus ends and less dynamic minus ends. Microtubules are required for vital processes in eukaryotic cells including mitosis, meiosis, maintenance of cell shape and intracellular transport. Microtubules are also necessary for movement of cells by means of flagella and cilia. In mammalian tissue culture cells microtubules have their minus ends anchored in microtubule organizing centers (MTOCs).The GTP (guanosintriphosphate) molecule is an essential for tubulin heterodimer to associate with other heterodimers to form microtubule. In vivo, microtubule dynamics vary considerably. Microtubule polymerization is reversible and a populations of microtubules in cells are on their minus ends either growing or shortening – this phenomenon is called dynamic instability of microtubules. On a practical level, microtubules can easily be stabilized by the addition of non-hydrolysable analogues of GTP (eg. GMPPCP) or more commonly by anti-cancer drugs such as Taxol. Taxol stabilizes microtubules at room temperature for many hours. Using limited proteolysis by enzymes both tubulin subunits can be divided into N-terminal and C-terminal structural domains. The alpha-tubulin (relative molecular weight around 50 kDa) is globular protein that exists in cells as part of soluble alpha/beta-tubulin dimer or it is polymerized into microtubules. In different species it is coded by multiple tubulin genes that form tubulin classes (in human 6 genes). Expressed tubulin genes are named tubulin isotypes. Some of the tubulin isotypes are expressed ubiquitously, while some have more restricted tissue expression. Alpha-tubulin is also subject of numerous post-translational modifications. Tubulin isotypes and their posttranslational modifications are responsible for multiple tubulin charge variants - tubulin isoforms. Heterogeneity of alpha-tubulin is concentrated in C-terminal structural domain. The beta-tubulin (relative molecular weight around 50 kDa) is counterpart of alpha-tubulin in tubulin heterodimer, it is coded by multiple tubulin genes and it is also posttranslationally modified. Heterogeneity of subunit is concentrated in C-terminal structural domain. |
| References: |
*Linhartova I, Draberova E, Viklicky V, Draber P.: Distribution of non-class-III beta-tubulin isoforms in neuronal and non-neuronal cells. FEBS Lett. 1993 Mar 29;320(1):79-82.
*Cowan NJ, Dobner PR, Fuchs EV, Cleveland D.W.: Expression of human alpha-tubulin genes: interspecies conservation of 3' untranslated regions.Mol Cell Biol. 1983 Oct;3(10):1738-45.
*Draberova E, Draber P: Novel monoclonal antibodies TU-08 and TU-16 specific for tubulin subunits. Folia Biol (Praha). 1998;44(1):35-6.
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