Proteins

Since 1993 Cytoskeleton has provided purified tubulin proteins to the scientific community. We continually strive to provide the purest, most biological active and relevant tubulin proteins, kits and reagents for today's researchers. The available tubulin proteins are highly purified and contain high biological activity. There are tubulins available from different species as well as different tissues within the same species. For general research purposes we recommend >99% pure tubulin from brain tissue e.g. Cat. # T240. This is a highly versatile substrate for microtubule binding studies as well as microtubule polymerization assays. Other more specialized tubulins include those from cancer cells which is used for drug screening and those from plant or fungal sources which are suitable for studies in their respective areas. Detailed information about each product can be found by clicking on the datasheet icon in the table below.

 

Cytoskeleton also provides many of these proteins in ready to use kit format, if you don't see the kit that fills your need please contact tservice@cytoskeleton.com for more assistance.

Cytoskeleton's products have been cited hundreds of times over the past 18 years. A select few are described here, for more citations on individual products please use the "Citations" tab on each individual product page.

Pure Tubulins, e.g. Tubulin protein (>99% pure): porcine brain (Cat. # T240)

Barten et al., 2011.  Tau transgenic mice as models for cerebrospinal fluid tau biomarkers.  J. Alzheimers Dis. 24, 1-15.   

Iuchi et al., 2009.  Heterocyclic Organobismuth(III) Compound Targets Tubulin to Induce G2/M Arrest in HeLa Cells.  J. Pharmacol. Sci. 109, 573-582.

Sugiyama et al., 2009.  Quick Shear-Flow Alignment of Biological Filaments for X-ray Fiber Diffraction Facilitated by Methylcellulose. Biophys. J. 97, 3132-3138.

Ishii et al., 2010. Image analysis of α/β-tubulin rings in two-dimensional crystalline arrays of periodic mesoporous nanostructures. J. Biochem. 147, 555-563.

MAP-enriched tubulins (70% tubulin + 30% MAPs): porcine brain (Cat. # ML116)

Tolg et al., 2010. RHAMM Promotes Interphase Microtubule Instability and Mitotic Spindle Integrity through MEK1/ERK1/2 Activity. J. Biol. Chem. 285, 26461-26474.

Cancer cell tubulin, e.g. Tubulin protein: Hela cancer cell (Cat. # H001)

Meng et al., 2008. A novel class of tubulin inhibitors that exhibit potent antiproliferation and in vitro vessel-disrupting activity. Cancer Chemother. Pharmacol. 61, 953-963.

Nunes, M., Kaplan, J., Wooters, J., Hari, M., Minnick, A. A., Jr., May, M. K., Shi, C., Musto, S., Beyer, C., Krishnamurthy, G. et al. (2005). Two photoaffinity analogues of the tripeptide, hemiasterlin, exclusively label α-tubulin. Biochemistry 44, 6844-6857.

Spittle, C., Charrasse, S., Larroque, C. and Cassimeris, L. (2000). The interaction of TOGp with microtubules and tubulin. J. Biol. Chem. 275, 20748-20753.

Fluorescent conjugates, e.g. Tubulin protein (X-rhodamine labeled): bovine brain (Cat. # TL620M)

Jimenez et al., 2011. Towards high throughput production of artificial egg oocytes using microfluidics. Lab Chip. 11, 429-434.

Cross and Powers, 2011. Nup98 regulates bipolar spindle assembly through association with microtubules and opposition of MCAK. Mol. Biol. Cell. 22, 661-672.

Biotin conjugates, e.g. Tubulin protein (biotin): porcine brain (Cat. # T333P).

Dixit and Ross (2010). Chapter 27-Studying plus-end tracking at single molecule resolution using TIRF microscopy. Methods in Cell Biology 95 ,543-554.

Gell et al. (2010). Chapter 13-Microtubule Dynamics Reconstituted In Vitro and Imaged by Single-Molecule Fluorescence Microscopy. Methods in Cell Biology 95 ,221-245.

Tubulin protein (>99% pure): porcine brain (Cat. # T238P)

King, S. J. and Schroer, T. A. (2000). Dynactin increases the processivity of the cytoplasmic dynein motor. Nat. Cell Biol. 2, 20-24.

Hong, M., Zhukareva, V., Vogelsberg-Ragaglia, V., Wszolek, Z., Reed, L., Miller, B. I., Geschwind, D. H., Bird, T. D., McKeel, D., Goate, A. et al. (1998). Mutation-specific functional impairments in distinct tau isoforms of hereditary FTDP-17. Science 282, 1914-1917.

Melander Gradin, H., Larsson, N., Marklund, U. and Gullberg, M. (1998). Regulation of microtubule dynamics by extracellular signals: cAMP-dependent protein kinase switches off the activity of oncoprotein 18 in intact cells. J. Cell Biol. 140, 131-141.

Sosa, H., Dias, D. P., Hoenger, A., Whittaker, M., Wilson-Kubalek, E., Sablin, E., Fletterick, R. J., Vale, R. D. and Milligan, R. A. (1997). A model for the microtubule-Ncd motor protein complex obtained by cryo-electron microscopy and image analysis. Cell 90, 217-224.

Pre-formed microtubules (Cat. # MT001 and MT002)

Pizzi et al., 2010. Evidences of new biophysical properties of microtubules. In: Artificial Neural Networks, Ed. S.J. Kwon, Nova Science Publishers Inc. Chapter 20.

Zhang et al., 2005. Development of a high-throughput robotic fluorescence-based assay for HsEg5 inhibitor screening. Anal. Biochem. 345, 326-335.

Question 1:  What tubulin product do you recommend for first-time users?   

Answer 1: There are multiple factors that influence tubulin stability and polymerization, including the presence and concentration of GTP, magnesium ions and glycerol, as well as temperature and tubulin concentrations.  To help the first-time tubulin user navigate these potential pitfalls, we recommend using our 99% pure tubulin from porcine brain (Cat. # T240 or BK011P) or for small quantities use the Microtubules/Tubulin Biochem Kit (Cat. # BK015).  The purified tubulin is ideal for polymerization assays designed to evaluate the efficacy (Vmax, IC50, EC50) of anti-cancer and anti-microtubule drugs and the datasheet (Cat. # T240) provides detailed instructions (and background information) on setting up resuspending, storing and polymerizing the tubulin.  The Microtubules/Tubulin Biochem Kit (Cat. # BK015) provides the researcher with the necessary reagents and instructions (and background information) to reproducibly prepare microtubules of a predetermined mean length.  After polymerization, the microtubules can be used immediately or stabilized with taxol before use, depending upon the needs of the experimental system.  The microtubules are ideal for motility experiments or microtubule-associated protein (MAP) binding assays.  The kit contains 99% pure tubulin (Cat. # TL238-A), general tubulin buffer (Cat. # BST01-001), tubulin glycerol buffer (Cat. # BST05-001), GTP stock (Cat. # BST06-001), taxol stock (Cat. # TXD01) and DMSO.

  

Question 2:  If the molecular weight of tubulin is 110,000 daltons why does it run at 50-55 kDa on a SDS-PAGE gel?

Answer 2: Under normal physiological conditions, monomeric tubulin consists of a heterodimer of one alpha and one beta isotype and each tubulin isotype is 55 kDa in size.  SDS-PAGE analysis denatures the heterodimer and shows tubulin running as a 55 kDa species.  Both alpha and beta isoforms are found at 55 kDa, since the 110 kDa heterodimer has been denatured.  Typically, the molar equivalent of tubulin is defined as the heterodimer which has a molecular weight of 110 kDa.

 

Question 3:  I need tubulin isolated and purified from my cell line.  How can I do that?

Answer 3: Cytoskeleton offers custom protein purification services and we are experts at isolating and purifying tubulin from multiple sources, including mammalian brains and HeLa and MCF-7 cancer cell lines.  To read a paper that utilized our cancer tubulins, please see Meng et al., 2008 (A novel class of tubulin inhibitors that exhibit potent antiproliferation and in vitro vessel-disrupting activity. Cancer Chemother. Pharmacol. 61, 953-963).  Please contact Technical Support at tservice@cytoskeleton.com to discuss your specific tubulin needs regarding isolation and purification from specific cell lines or tissue samples.

 

   If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com.

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