Western Blotting (WB), Immunocytochemistry (ICC), Flow Cytometry (FACS)
Specificity
The antibody TU-30 recognizes C-terminus (amino acids 434-449 in human) of gamma-tubulin, a 48 kDa structural constituent of cytoskeleton and microtubule organizing center (MTOC). The epitope was located in the amino acid sequence TRPDYI (aa439-444 in human), which is present on human gamma-tubulin 1 but not on human gamma-tubulin 2.
Cross-Reactivity (Details)
Human, Porcine, Mouse, Rat, Bovine, Chicken, Protozoa, Plants
Purification
Purified by protein-A affinity chromatography.
Purity
> 95 % (by SDS-PAGE)
Immunogen
C-terminal peptide of gamma-tubulin counjugated to KLH.
Immunocytochemistry: Recommended dilution: 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. Positive control: P-19 murine embryonal carcinoma cell line, 3T3 murine fibroblasts. The antibody TU-30 stains only fixed cells. Western blotting: Recommended dilution 1-2 μg/mL, reducing conditions.
Restrictions
For Research Use only
Concentration
1 mg/mL
Buffer
Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Preservative
Sodium azide
Precaution of Use
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Do not freeze.
Storage
4 °C
Storage Comment
Store at 2-8°C. Do not freeze.
Vulprecht, David, Tibelius, Castiel, Konotop, Liu, Bestvater, Raab, Zentgraf, Izraeli, Krämer: "STIL is required for centriole duplication in human cells." in: Journal of cell science, (2012) (PubMed).
Löffler, Fechter, Matuszewska, Saffrich, Mistrik, Marhold, Hornung, Westermann, Bartek, Krämer: "Cep63 recruits Cdk1 to the centrosome: implications for regulation of mitotic entry, centrosome amplification, and genome maintenance." in: Cancer research, Vol. 71, Issue 6, pp. 2129-39, (2011) (PubMed).
Koledova, Kafkova, Kraemer, Divoky: "DNA damage-induced degradation of Cdc25A does not lead to inhibition of Cdk2 activity in mouse embryonic stem cells." in: Stem cells (Dayton, Ohio), Vol. 28, Issue 3, pp. 450-61, (2010) (PubMed).
Katsetos, Reddy, Dráberová, Smejkalová, Del Valle, Ashraf, Tadevosyan, Yelin, Maraziotis, Mishra, Mörk, Legido, Nissanov, Baas, de Chadarévian, Dráber: "Altered cellular distribution and subcellular sorting of gamma-tubulin in diffuse astrocytic gliomas and human glioblastoma cell lines." in: Journal of neuropathology and experimental neurology, Vol. 65, Issue 5, pp. 465-77, (2006) (PubMed).
Sulimenko, Dráberová, Sulimenko, Macurek, Richterová, Dráber, Dráber: "Regulation of microtubule formation in activated mast cells by complexes of gamma-tubulin with Fyn and Syk kinases." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 176, Issue 12, pp. 7243-53, (2006) (PubMed).
Libusová, Sulimenko, Sulimenko, Hozák, Dráber: "gamma-Tubulin in Leishmania: cell cycle-dependent changes in subcellular localization and heterogeneity of its isoforms." in: Experimental cell research, Vol. 295, Issue 2, pp. 375-86, (2004) (PubMed).
Binarová, Cenklová, Hause, Kubátová, Lysák, Dolezel, Bögre, Dráber: "Nuclear gamma-tubulin during acentriolar plant mitosis." in: The Plant cell, Vol. 12, Issue 3, pp. 433-42, (2000) (PubMed).
Nováková, Dráberová, Schürmann, Czihak, Viklický, Dr-aber: "gamma-Tubulin redistribution in taxol-treated mitotic cells probed by monoclonal antibodies." in: Cell motility and the cytoskeleton, Vol. 33, Issue 1, pp. 38-51, (1996) (PubMed).
Tubulin gamma 1,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. Humans possess two gamma-tubulin genes. Gamma-tubulin 1 represents a ubiquitous isotype, whereas gamma-tubulin 2 is found predominantly in the brain, where it may be endowed with divergent functions beyond microtubule nucleation.,TUBG