Best-in-Class Tools to Accelerate Your Tubulin Discoveries

Cytoskeleton Inc. has been at the forefront of tubulin research since 1993, providing biologically active tubulin proteins, kits, and reagents to the scientific community.

Through our collaboration, Tebubio has brought these high-quality products to researchers across Europe, helping to unlock the complex dynamics of the cytoskeleton.

Our tubulin-related products and functional assays are designed to:

• Investigate tubulin dynamics

• Identify binding partners

• Analyse kinetic properties

All while saving researchers valuable time and enhancing scientific productivity.

Labelled Tubulin Examples (Fig. 1):

• HiLyte Fluor™ 488 Labelled Tubulin

• TRITC Rhodamine Labelled Tubulin

For researchers setting up polymerisation assays or conducting drug screenings, bulk quantities of tubulin preparations are available upon request.

Please find here an overview of the available labelled. 

Figure 1: HiLyte Fluor™ 488 Labelled Tubulin (left) and TRITC Rhodamine Labelled Tubulin (right).

Figure 2: Microtubule Schematic.

To provide direct experimental access to the filamentous form of tubulin, we can also offer pre-made microtubules (Fig. 2)  e.g. as substrates for kinesin motor proteins. Kinesin motor proteins orchestrate a wide range of kinetic events within a cell. They have been shown to move cargoes such as chromosomes and vesicles along Microtubule tracks.

Cytoskeleton’s pre-formed Microtubules are an excellent substrate for detecting microtubule-binding proteins in combination with the Kinesin Enzyme Linked Inorganic Phosphate ATPase Kit.

Tubulin Polymerisation Assays (Fig. 3):

• Fluorescence-based: highly sensitive, cost-effective, suitable for high-throughput studies

• Absorbance-based: classical method widely used in tubulin research
  Applications include drug screening, IC50 determination, protein effect studies, and educational purposes.

Figure 3: Tubulin polymerisation using the fluorescence-based tubulin polymerisation assay. Tubulin was incubated alone (Control), with Paclitaxel or Vinblastine. Each condition was tested in duplicate. Polymerisation was measured by excitation at 360 nm and emission at 420 nm.  The three Phases of tubulin polymerisation are marked for the control polymerisation curve: I: nucleation, II: growth, III: steady state equilibrium.

In Vivo Microtubules/Tubulin Assay (Fig. 4):

• Quantifies cellular microtubule versus free-tubulin content

• Ideal for studying drug effects, mutant cell lines, or environmental influences on the cytoskeleton

Figure 4: WB analysis of low speed (LS) and ultra-centrifuged (HS) samples of untreated and paclitaxel-treated tissue culture cells. Untreated or Taxol-treated (1 µM Taxol,1h, 37 °C, 5%CO2) 3T3 cell lysates were centrifuged at low speed (1,000 g), and the low-speed pellet was saved for western analysis (LSP). The low-speed supernatants were centrifuged at 100,000 g, and both supernatant (HSS) and pellet (HSP) were saved for western blot analysis. All lanes were loaded with cell lysate that represents an equal number of cells per lane. Western blots were probed with anti-tubulin antibody, and tubulin signals were quantified by densitometry. The majority of taxol-stabilised microtubules appear in the low-speed pellet (LSP/Taxol treated), whereas the majority of microtubules from the untreated samples appear in the high-speed supernatant. The difference in microtubule fractionation reflects the stabilisation of taxol-treated microtubules in 3T3 cells.