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Issue 3
Due to its essential role in cell division, the mitotic spindle has become an important therapeutic target for
anticancer agents such as the Vinca alkaloids or taxanes. However, various mechanisms of developed
resistance cause that these drugs often fail to induce desired mitotic block and death of cancer cells. Further
investigation of microtubular system, namely full understanding its assembly, dynamics and regulation,
is thus an incentive goal both for clinical and basic researchers. (1–3)
Microtubules of mitotic spindle
(green) originate from γ-tubulin
rings (red) localized to pericentriolar
material of centrosomes
In animal cells, a center of microtubule organization is the centrosome composed of a pair of cylindrical
centrioles surrounded by fibrous pericentriolar material. Microtubules are nucleated from γ-tubulin ring
structures embedded in the pericentriolar material. Formation of mitotic spindle is preceded by duplication of
centrosome during S phase. Before mitosis, both centrosomes increase their microtubule nucleation capacity
and form two microtubule asters that are pushed apart from each other by the forces of motor proteins associated
at the microtubule surface. Upon nuclear envelope breakdown, formation of spindle apparatus is finalized
by binding chromosomal kinetochores to centrosomal microtubules. (4, 5)
Exbio offers the excellent TU-30 monoclonal antibody that recognizes C-terminus of γ-tubulin. This
antibody decorates interphase in mammalian cell centrosomes as well as half-spindles and midbodies
in mitotic cells of various origin. (6–10)
Clone
Isotype (Host)
Reactivity
Application
Catalogue Nr.
Format
TU-30
IgG2b (Mouse)
Human, Porcine,
Mouse, Rat, Chicken,
Protozoa, Plants
Immunocytochemistry,
Western Blotting
11-465-C100
11-465-M001
1C-465-C100
1D-465-C100
Purified (0.1 mg)
Purified (1.0 mg)
DY547 (0.1 mg)
DY647 (0.1 mg)
EXBIO Praha, a.s. | Nad Safinou II 366 | 252 42 Vestec | Czech Republic | tel.: +420 261 090 666 | fax: +420 261 090 660
e-mail: [email protected] | www.exbio.cz
Cytoskeleton
Detection of Mitotic Spindle
A
B
D
E
C
F
DY547
Trimethine cyanine
(Spectrally similar to Cy3, TRITC, Alexa 546, Alexa 555)
Absorption / Emission max.:
557 nm / 574 nm
Molar a bsorbance:
150.000 M-1cm-1
Zhou J. and Giannakakou P.; Curr. Med. Chem. Anticancer Agents 2005; 5: 65-71 (1)
Verrills N.M. and Kavallaris M.; Curr. Pharm. Des. 2005; 11: 1719-1733 (2)
Liaw T.Y. et al.; Curr. Drug Targets 2007; 8: 739-749 (3)
Wiese C. and Y. Zheng; J. Cell Sci. 2006; 119: 4143-4153 (4)
Haren L. et al.; J. Cell Biol. 172: 505-515 (5)
DY647
Pentamethine-based fluorophore
(Spectrally similar to Cy5, Alexa 647)
Absorption / Emission max.:
653 nm / 672 nm
Molar a bsorbance:
250.000 M-1cm-1
Novakova M. et al.; Cell Motil. Cytoskeleton 1996; 33: 38-51 (6)
Binarova P. et al.; Plant Cell 2000; 12: 433-442 (7)
Nohynkova E. et al.; Eur. J. Cell Biol. 2000; 79: 438-445 (8)
Linhartova I. et al.; Dev. Dyn. 2002; 223: 229-240 (9)
Libusova L. et al.; Exp. Cell Res. 2004; 295: 672-679 (10)
Distributed by/Contacts:
www.exbio.cz
DYO M I C S
A–C)
β-tubulin (green) were stained with Hoechst (DNA, blue) and centrosomes were visualized
Rat prostate carcinoma cells expressing GFP-β
using DY547-conjugated anti-γγ-tubulin monoclonal antibody TU-30 (red).
D–F)
α-tubulin
Mitotic spindle in 3T3 mouse fibroblasts (D – metaphase, E – anaphase, F – telophase) as detected by TU-30 anti-γγ-tubulin (red), anti-α
polyclonal (green) and DAPI (DNA, blue).