Tubulin protein (>99% pure): porcine brain

Tubulin protein (>99% pure): porcine brain

Product Uses Include

  • IC50 & EC50 determinations for anti-tubulin ligands.
  • Microtubule binding studies
  • Tubulin monomer binding studies
  • HDAC6 studies
  • Microtubule activated kinesin ATPase assays

Tubulin protein has been purified from porcine brain by an adaptation of the method of Shelanski et al. (1), Further purification to >99% purity was achieved by cation exchange chromatography. Tubulin is supplied as a white lyophilized powder.

Fully active for polymerization, this product is lyophilized with a patented technology for increased stability and longevity. T240 is stable for 1 year at 4°C desiccated. If your project requires the same batch of tubulin for consistent results, it is highly recommended that the item is purchased in bulk in order to save time and money. This product can be used as a substitute for our highly purified bovine tubulin products (Cat. # TL238, T238 and T237) and behaves in an identical fashion.

Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are >99% pure


Figure 1: A 20 µg sample of T240 protein was separated by electrophoresis 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 tubulin is defined as 5.0 mg of purified protein (as determined by the Precision Red Advanced Protein Assay Reagent cat. # ADV02). The biological activity of T240 is assessed by a tubulin polymerization assay. The ability of tubulin to polymerize into microtubules can be followed by observing an increase in optical density of the tubulin solution at 340 nm. A 5 mg/ml tubulin solution in General Tubulin Buffer buffer plus 5% glycerol and 1 mM GTP should achieve an OD340 nm reading between 0.75-1.10 in 30 min at 37°C when using a spectrophotometer pathlength of 0.8 cm (180 µl sample volume in a 1/2 area 96-well plate).

It should be noted that tubulin minus glycerol WILL NOT polymerize in G-PEM buffer until very high tubulin concentrations (>10 mg/ml). Even at these concentrations polymerization is comparatively slow. Efficient polymerization at low concentration of tubulin minus glycerol can be achieved by addition of a polymerization stimulating compound, e.g., glycerol, paclitaxel or DMSO.


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

For product Datasheets and MSDSs please click on the PDF links below.   For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

AuthorTitleJournalYearArticle Link
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Henty-Ridilla, Jessica L.Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence (TIRF) MicroscopyJoVE (Journal of Visualized Experiments)2022ISSN 1940--087X
Sasanpour, Mehrzad et al.Reconstituting and Characterizing Actin-Microtubule Composites with Tunable Motor-Driven Dynamics and MechanicsJoVE (Journal of Visualized Experiments)2022ISSN 1940--087X
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Kuzmić, Mira et al.Septin-microtubule association via a motif unique to isoform 1 of septin 9 tunes stress fibersJournal of Cell Science2022ISSN 1477-9137
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Wu, R. et al.Thermal hysteresis in microtubule assembly/disassembly dynamics: The aging-induced degradation of tubulin dimersBiochemistry and Biophysics Reports2022ISSN 2405-5808
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Watson, Joseph L. et al.High-efficacy subcellular micropatterning of proteins using fibrinogen anchorsJournal of Cell Biology2021ISSN 1540-8140
Elahi, Montasir et al.High-fat diet–induced activation of SGK1 promotes Alzheimer’s disease–associated tau pathologyHuman Molecular Genetics2021ISSN 0964--6906
Zhou, Libin et al.Rare CASP6N73T variant associated with hippocampal volume exhibits decreased proteolytic activity, synaptic transmission defect, and neurodegenerationScientific Reports 2021 11:12021ISSN 2045--2322
Budaitis, Breane G. et al.Pathogenic mutations in the kinesin-3 motor KIF1A diminish force generation and movement through allosteric mechanismsJournal of Cell Biology2021ISSN 1540-8140
Habicht, Juri et al.UNC-45A breaks the microtubule lattice independently of its effects on non-muscle myosin IIJournal of Cell Science2021ISSN 1477-9137
Gao, Li et al.A Robust, GFP-Orthogonal Photoswitchable Inhibitor Scaffold Extends Optical Control over the Microtubule CytoskeletonCell Chemical Biology2021ISSN 2451-9448
Nakayama, Shogo et al.Planar cell polarity induces local microtubule bundling for coordinated ciliary beatingJournal of Cell Biology2021ISSN 1540-8140
Ludzia, Patryk et al.Structural characterization of KKT4, an unconventional microtubule-binding kinetochore proteinStructure2021ISSN 1878-4186
Kundu, Tanushree et al.Coupling of dynamic microtubules to F-actin by Fmn2 regulates chemotaxis of neuronal growth conesJournal of Cell Science2021ISSN 1477-9137
Yano, Tomoki et al.A microtubule‐LUZP1 association around tight junction promotes epithelial cell apical constrictionThe EMBO Journal2021ISSN 0261--4189
Tang, Qing et al.NDST3 deacetylates α‐tubulin and suppresses V‐ATPase assembly and lysosomal acidificationThe EMBO Journal2021ISSN 0261--4189
Yu, Xian et al.MARK4 controls ischaemic heart failure through microtubule detyrosinationNature2021ISSN 1476-4687
Alfieri, Angus et al.Two modes of PRC1-mediated mechanical resistance to kinesin-driven microtubule network disruptionCurrent Biology2021ISSN 1879-0445
Nourbakhsh, Kimya et al.TAOK2 is an ER-localized kinase that catalyzes the dynamic tethering of ER to microtubulesDevelopmental Cell2021ISSN 1878-1551
Kaur, Simranpreet et al.Expansion of the phenotypic spectrum of de novo missense variants in kinesin family member 1A (KIF1A)Human Mutation2020ISSN 1098-1004
Baker, Stacey J. et al.A Contaminant Impurity, Not Rigosertib, Is a Tubulin Binding AgentMolecular Cell2020ISSN 1097-4164
Ricketts, Shea N. et al.Triggering Cation-Induced Contraction of Cytoskeleton Networks via MicrofluidicsFrontiers in Physics2020ISSN 2296-424X
Diwaker, Drishya et al.Deletion of the Pseudorabies virus gE/gI-US9p complex disrupts kinesin KIF1A and KIF5C recruitment during egress, and alters the properties of microtubule-dependent transport in vitroPLoS Pathogens2020ISSN 1553-7374
Ouyang, Changhan et al.Autophagic degradation of KAT2A/GCN5 promotes directional migration of vascular smooth muscle cells by reducing TUBA/α-tubulin acetylationAutophagy2020ISSN 1554-8635
Müller-Deku, Adrian et al.Photoswitchable pac*******-based microtubule stabilisers allow optical control over the microtubule cytoskeletonNature Communications2020ISSN 2041-1723
Alatrash, Nagham et al.Disruption of microtubule function in cultured human cells by a cytotoxic ruthenium(ii) polypyridyl complexChemical Science2020ISSN 2041-6539
Kraus, Yvonne et al.Isoquinoline-based biaryls as a robust scaffold for microtubule inhibitorsEuropean Journal of Medicinal Chemistry2020ISSN 1768-3254
Aher, Amol et al.CLASP Mediates Microtubule Repair by Restricting Lattice Damage and Regulating Tubulin IncorporationCurrent Biology2020ISSN 1879-0445
Gunji, Shizuka et al.Excess Pyrophosphate Restrains Pavement Cell Morphogenesis and Alters Organ Flatness in Arabidopsis thalianaFrontiers in Plant Science2020ISSN 1664-462X
Martinez, Pablo et al.TANGLED1 mediates microtubule interactions that may promote division plane positioning in maizeJournal of Cell Biology2020ISSN 1540-8140
Chen, Keyu et al.Giant ankyrin-B suppresses stochastic collateral axon branching through direct interaction with microtubulesJournal of Cell Biology2020ISSN 1540-8140
Rodríguez-García, Ruddi et al.Mechanisms of Motor-Independent Membrane Remodeling Driven by Dynamic MicrotubulesCurrent Biology2020ISSN 1879-0445
Atherton, Joseph et al.The mechanism of kinesin inhibition by kinesin binding proteineLife2020ISSN 2050-084X
Saper, Gadiel et al.Kinesin-propelled label-free microtubules imaged with interference reflection microscopyNew Journal of Physics2020ISSN 1367-2630
Adriaans, Ingrid E. et al.MKLP2 Is a Motile Kinesin that Transports the Chromosomal Passenger Complex during AnaphaseCurrent Biology2020ISSN 1879-0445
Gaska, Ignas et al.The Mitotic Crosslinking Protein PRC1 Acts Like a Mechanical Dashpot to Resist Microtubule SlidingDevelopmental Cell2020ISSN 1878-1551
Francis, Madison L. et al.Non-monotonic dependence of stiffness on actin crosslinking in cytoskeleton compositesSoft Matter2019ISSN 1744-6848
Leong, Su Ling et al.Reconstitution of Microtubule Nucleation In Vitro Reveals Novel Roles for Mzt1Current Biology2019ISSN 0960-9822
Lopes, Joseph et al.Membrane mediated motor kinetics in microtubule gliding assaysScientific Reports2019ISSN 2045-2322
Nakos, Konstantinos et al.Regulation of microtubule plus end dynamics by septin 9Cytoskeleton2019ISSN 1949-3592
Ng, Cai Tong et al.Electron cryotomography analysis of Dam1C/DASH at the kinetochore-spindle interface in situJournal of Cell Biology2019ISSN 1540-8140
Guzman-Sepulveda, Jose Rafael et al.Tubulin Polarizability in Aqueous SuspensionsACS Omega2019ISSN 2470-1343
Zhang, Shengnan et al.In-cell NMR study of Tau and MARK2 phosphorylated TauInternational Journal of Molecular Sciences2019ISSN 1422-0067
Jiang, Shuo et al.Interplay between the Kinesin and Tubulin Mechanochemical Cycles Underlies Microtubule Tip Tracking by the Non-motile Ciliary Kinesin Kif7Developmental Cell2019ISSN 1878-1551
Karasmanis, Eva P. et al.Erratum: Polarity of Neuronal Membrane Traffic Requires Sorting of Kinesin Motor Cargo during Entry into Dendrites by a Microtubule-Associated Septin (Developmental Cell (2018) 46(2) (204–218.e7), (S1534580718304982) (10.1016/j.devcel.2018.06.013))Developmental Cell2018ISSN 1878-1551
Ganguly, Anindya et al.Importin-β Directly Regulates the Motor Activity and Turnover of a Kinesin-4Developmental Cell2018ISSN 1878-1551
Fan, Yuanwei et al.The Arabidopsis SPIRAL2 Protein Targets and Stabilizes Microtubule Minus EndsCurrent Biology2018ISSN 0960-9822
Romé, Pierre et al.A novel microtubule nucleation pathway for meiotic spindle assembly in oocytesJournal of Cell Biology2018ISSN 1540-8140
Tripathy, Ratna et al.Mutations in MAST1 Cause Mega-Corpus-Callosum Syndrome with Cerebellar Hypoplasia and Cortical MalformationsNeuron2018ISSN 1097-4199
Rao, Lu et al.Combining structure–function and single-molecule studies on cytoplasmic dyneinMethods in Molecular Biology2018ISSN 1064-3745
McClintock, Mark A. et al.RNA-directed activation of cytoplasmic dynein-1 in reconstituted transport RNPseLife2018ISSN 2050-084X
Murray, John W. et al.Reduction of organelle motility by removal of potassium and other solutesPLoS ONE2017ISSN 1932-6203
Kandel, Mikhail E. et al.Label-Free Imaging of Single Microtubule Dynamics Using Spatial Light Interference MicroscopyACS Nano2017ISSN 1936-086X
Okeyoshi, Kosuke et al.Methods for the self-integration of megamolecular biopolymers on the drying air-LC interfaceJournal of Visualized Experiments2017ISSN 1940-087X
Majellaro, Maria et al.Investigating Structural Requirements for the Antiproliferative Activity of Biphenyl NicotinamidesChemMedChem2017ISSN 1860-7187
Onder, Seda et al.Monoclonal Antibody That Recognizes Diethoxyphosphotyrosine-Modified Proteins and Peptides Independent of Surrounding Amino AcidsChemical Research in Toxicology2017ISSN 1520-5010
Monda, Julie K. et al.Microtubule Tip Tracking by the Spindle and Kinetochore Protein Ska1 Requires Diverse Tubulin-Interacting SurfacesCurrent Biology2017ISSN 0960-9822
Howes, Stuart C. et al.Structural differences between yeast and mammalian microtubules revealed by cryo-EMJournal of Cell Biology2017ISSN 1540-8140
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Matsuyama, Tomonori et al.Midaz(olam) inhibits the hypoxia-induced up-regulation of erythropoietin in the central nervous systemEuropean Journal of Pharmacology2015ISSN 1879-0712
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Question 1:  What is the proper way to store the tubulin to insure maximum stability and activity?

Answer 1:  The recommended storage condition for the lyophilized tubulin product is 4°C with desiccant to maintain humidity at <10% humidity.  Under these conditions the protein is stable for 6 months.  Lyophilized protein can also be stored desiccated at -70°C where it will be stable for 6 months.  However, at -70°C the rubber seal in the lid of the tube could crack and allow in moisture.  Therefore we recommend storing at 4°C.  If stored at -70°C, it is imperative to include desiccant with the lyophilized protein if this storage condition is utilized.  After reconstituting the protein as directed, the concentrated protein in G-PEM buffer should be aliquoted, snap frozen in liquid nitrogen and stored at -70°C (stable for 6 months).  NOTE: It is very important to snap freeze the tubulin in liquid nitrogen as other methods of freezing will result in significantly reduced activity.  Defrost rapidly by placing in a room temperature water bath for 1 min.  Avoid repeated freeze/thaw cycles.


Question 2:  Why does Cytoskeleton recommend the use of general tubulin buffer and GTP for resuspending tubulin?

Answer 2:  We recommend resuspending tubulin in general tubulin buffer + GTP to maintain tubulin monomer protein stability and conformation and to provide the necessary components for polymerization.  For resuspension, we recommend using a general tubulin buffer (Cat. # BST01-001) which consists of 80 mM PIPES, 2 mM MgCl2, 1 mM EGTA, pH 7.0, supplemented with 1 mM GTP (Cat. # BST06-001).  Tubulin requires GTP and magnesium ions for proper stability and conformation, even in its monomeric state.  GTP is also required for the polymerization process as its hydrolysis during tubulin polymerization is necessary for polymerization to occur.  EGTA is a chelator of calcium which is a potent inhibitor of tubulin polymerization.  Glycerol is often added to a final concentration of 5 - 10% to enhance polymerization; however, glycerol is not necessary for the maintenance of biologically active tubulin and does not need to be included when reconstituting and storing tubulin.  When aliquoting reconstituted tubulin for storage, it is essential to aliquot and snap-freeze tubulin in liquid nitrogen at a concentration of >6 mg/ml to preserve tubulin’s biological activity.  Then the aliquots should be stored at -70°C.  When thawing the aliquots, thaw rapidly in a room temperature water bath and place on ice until right before experimental use.


Question 3: How does porcine tubulin compare to bovine tubulin?

Answer 3: Click here for an in-depth comparison of porcine and bovine tubulin.

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