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Mouse Monoclonal to gamma-tubulin

TU-30 (IgG1)

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Background

The gamma-tubulin (TUBG1; relative molecular weight about 48 kDa) is a minor member of tubulin family (less that 0.01% of tubulin dimer). The gamma-tubulin ring structures, however, serve to provide structural primer for initiation of microtubular nucleation and growth, thereby being crutial for microtubule-based cellular processes, above all for mitotic spindle formation. In animal cells, a center of microtubule organization is the centrosome composed of a pair of cylindrical centrioles surrounded by fibrous pericentriolar material containing gamma-tubulin. Formation of the mitotic spindle is preceded by duplication of centrosome during S phase. Before mitosis, both centrosomes increase their microtubule nucleation capacity and form two microtuble asters that are pushed apart from each other by the forces of motor proteins associated at the microtubule surface.

Specificity

The antibody TU-30 recognizes C-terminal peptide sequence EYHAATRPDYISWGTQ (aa 434-449) of gamma-tubulin, a 48 kDa structural constituent of cytoskeleton and microtubule organizing center (MTOC).

Regulatory Status

Immunogen

C-terminal peptide of gamma-tubulin counjugated to KLH.

Species Reactivity:

  • Human
  • Porcine
  • Mouse
  • Rat
  • Bovine
  • Chicken
  • Protozoa
  • Plants

Negative Species:

Applications:

  • Immunocytochemistry
    Recommended dilution:
    Purified Antibody: 1-2 μg/ml
    Staining technique: (a) Fix cells for 10 min in methanol at -20°C and for 6 min in acetone at -20°C; (b) Fix cells directly in methanol for 10 min at -20°C or in acetone for 10 min at -20°C.
    Incubation: 45 min in room temperature
    Positive control:
    P-19 mouse embryonal carcinoma cell line
    3T3 mouse fibroblasts
    Application note: The antibody TU-30 stains only fixed cells.
  • Western Blotting
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

  • *Wiese C, Zheng Y: Microtubule nucleation: gamma-tubulin and beyond. J Cell Sci. 2006 Oct 15;119(Pt 20):4143-53. [Abstract] [Full Text]
  • *Haren L, Remy MH, Bazin I, Callebaut I, Wright M, Merdes A. NEDD1-dependent recruitment of the gamma-tubulin ring complex to the centrosome is necessary for centriole duplication and spindle assembly. J Cell Biol. 2006 Feb 13;172(4):505-15. [Abstract] [Full Text]
  • *Draberova L, Draberova E, Surviladze Z, Draber P, Draber P: Protein tyrosine kinase p53/p56 (lyn) forms complexes with gamma-tubulin in rat basophilic leukemia cells. Int Immunol. 1999 Nov;11(11):1829-39. [Abstract] [Full Text]
  • Product Specific References

  • *Novakova M, Draberova E, Schurmann W, Czihak G, Viklicky V, Draber P: gamma-Tubulin redistribution in taxol-treated mitotic cells probed by monoclonal antibodies. Cell Motil Cytoskeleton. 1996;33(1):38-51. [Abstract]
  • *Binarova P, Cenklova V, Hause B, Kubatova E, Lysak M, Dolezel J, Bogre L, Draber P: Nuclear gamma-tubulin during acentriolar plant mitosis. Plant Cell. 2000 Mar;12(3):433-42. [Abstract] [Full Text]
  • *Libusova L, Sulimenko T, Sulimenko V, Hozak P, Draber P: gamma-Tubulin in Leishmania: cell cycle-dependent changes in subcellular localization and heterogeneity of its isoforms. Exp Cell Res. 2004 May 1;295(2):375-86. [Abstract]
  • *Katsetos CD, Reddy G, Dráberová E, Smejkalová B, Del Valle L, Ashraf Q, Tadevosyan A, Yelin K, Maraziotis T, Mishra OP, Mörk S, Legido A, Nissanov J, Baas PW, de Chadarévian JP, Dráber P: Altered cellular distribution and subcellular sorting of gamma-tubulin in diffuse astrocytic gliomas and human glioblastoma cell lines. J Neuropathol Exp Neurol. 2006 May;65(5):465-77. [Abstract]
  • *Sulimenko V, Dráberová E, Sulimenko T, Macurek L, Richterová V, Dráber P, Dráber P: Regulation of microtubule formation in activated mast cells by complexes of gamma-tubulin with Fyn and Syk kinases. J Immunol. 2006 Jun 15;176(12):7243-53. [Abstract] [Full Text]
  • For research use only. Not for drug, diagnostic or other use.

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  • Example Data



    TU-30
    Fig. 1 Immunofluorescence staining (mouse embryonal carcinoma cells)
    TU-30
    Fig. 2 Immunofluorescence staining (mouse fibroblasts)

    Fig. 1. Immunofluorescence staining of P19X1 mouse embryonal carcinoma cell line using anti-gamma-tubulin (TU-30) (detection by secondary antibody Goat anti-mouse Cy3). Nuclei were stained with DAPI (blue).

    Fig. 2. Immunofluorescence staining of mouse fibroblasts using anti-gamma-tubulin (TU-30; direct conjugate with Dyomics 547, red). Nuclei were stained with DAPI (blue).





    TU-30
    Fig. 3A, 3B, 3C Immunofluorescence staining (mouse fibroblasts)

    Fig. 3. Immunofluorescence staining of microtubular networks in 3T3 mouse fibroblasts.
    3A – metaphase; 3B – anaphase; 3C - telophase
    Gamma-tubulin (red) stained with anti-gamma-tubulin (TU-30), alpha-tubulin (green) with polyclonal anti-alpha-tubulin antibody and nuclei with DAPI (blue).









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