Tubulin protein: MAP rich: bovine brain

Studying the effects of MAPs on microtubule dynamics
Studying the binding sites between MAPs and tubulin.
Studying the effects of anti-microtubule drugs on microtubule polymerization.

Material
Tubulin and microtubule associated proteins (MAPs) has been purified from bovine brain by an adaptation of the method of Shelanski et al. (1). Tubulin is supplied as a white lyophilized powder. The protein composition is approximately 70% tubulin (55 kDa heterodimer) and 30% MAPs (see Figure 1). The MAPs in this product act to stabilize microtubules and to enhance tubulin polymerization. MAP rich tubulin can polymerize efficiently at 1-2 mg/ml.

MAP-rich tubulin is also available from a porcine source (Cat. # ML116)

Purity
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are approximately 70% tubulin and 30% MAPs.

	Figure 1: 100 µg of ML113 was run on a 10% SDS-PAGE gel and stained with Coomassie Blue. Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat. # ADV02).

Biological Activity
One unit of ML113 is defined as 5.0 mg of protein (as determined by the Precision Red Protein Assay Reagent (cat. # ADV02). MAP-rich tubulin will polymerize efficiently at protein concentrations above 1 mg/ml and temperatures above 24°C.

	Figure 2: MAP-rich tubulin polymerization under different conditions. Samples of ML113 in G-PEM buffer were placed in 96-well plates and incubated at the specified temperatures. Sample 1: 2 mg/ml at 37°C. Sample 2: 2 mg/ml at 24°C. Sample 3: 1 mg/ml at 24°C. Sample 4, same as 3 except in the presence of 3 µM nocodazole. Tubulin polymerization was observed by measuring the optical density at 340 nm over time.

References

Shelanski M. L., et al. (1973). Proc. Natl. Acad. Sci. USA. 70, 765-768

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Product Description

MSDS

Datasheet

Cat #

Tubulin protein: MAP rich: bovine brain ML113

Kuo, C. C., Hsieh, H. P., Pan, W. Y., Chen, C. P., Liou, J. P., Lee, S. J., Chang, Y. L., Chen, L. T., Chen, C. T. and Chang, J. Y. (2004). BPR0L075, a novel synthetic indole compound with antimitotic activity in human cancer cells, exerts effective antitumoral activity in vivo. Cancer Res. 64, 4621-4628.

Haggarty, S. J., Koeller, K. M., Wong, J. C., Grozinger, C. M. and Schreiber, S. L. (2003). Domain-selective small-molecule inhibitor of histone deacetylase 6 (HDAC6)-mediated tubulin deacetylation. Proc. Natl. Acad. Sci. U. S. A. 100, 4389-4394.

Xiao, D., Pinto, J. T., Soh, J. W., Deguchi, A., Gundersen, G. G., Palazzo, A. F., Yoon, J. T., Shirin, H. and Weinstein, I. B. (2003). Induction of apoptosis by the garlic-derived compound S-allylmercaptocysteine (SAMC) is associated with microtubule depolymerization and c-Jun NH2-terminal kinase 1 activation. Cancer Res. 63, 6825-6837.

Xie, Z., Sanada, K., Samuels, B. A., Shih, H. and Tsai, L. H. (2003). Serine 732 phosphorylation of FAK by Cdk5 is important for microtubule organization, nuclear movement, and neuronal migration. Cell 114, 469-482.

Wang, H., Bedford, F. K., Brandon, N. J., Moss, S. J. and Olsen, R. W. (1999). GABA(A)-receptor-associated protein links GABA(A) receptors and the cytoskeleton. Nature 397, 69-72.

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