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Microtubules have many amazing properties, not least of which being their dynamic behavior during the cell cycle. Regulation of microtubules involves multiple proteins and complexes; identifying the mechanisms that generate and control tubulin dynamics, and understanding how these affect microtubule function are long-standing goals in this field.
Popular assays include the use of specific antibodies in immunofluorescent imaging, and agents that modify different microtubule components. Here we provide the relevant tools to study these important microtubule processes.
|Microtubule nucleation||γ-tubulin||Forms part of the microtubule organizing centers (MTOCs), and facilitates tubulin dimer polymerization.||γ-tubulin mouse monoclonal antibody [GTU-88]|
|Microtubule polymerization||α-tubulin||Polymerizes with β-tubulin to form microtubules.|
|β-tubulin||Polymerizes with α-tubulin to form microtubules.|
|Microtubule depolymerization||Kinesins||The kinesin-13 class depolymerizes microtubule by bending tubulins at microtubule ends.|
Immunocytochemistry of formaldehyde-fixed HeLa cells stained with anti-beta tubulin antibody (ab6046). The secondary antibody (green) was ab150081 Alexa Fluor® 488 goat anti-rabbit IgG. Nuclear DNA was labeled with DAPI (shown in blue). The negative control is a secondary only assay. Click here to view the full datasheet for ab6046.
|Post-translational modifications (PTMs)||Acetylation||Acetylation on the K40 residue contributes to microtubule stability and cell motility. Deacetylation is linked to cell cycle progression and cancer.|
|Detyrosination||Involved in cell differentiation and microtubule stabilization.|
|Polyglutamylation||Occurs when glutamate side chains are formed on glutamate residues. Catalyzed by tubulin tyrosine ligase like (TTLL) enzymes.|
|Polyamination||Contributes to microtubule stabilization. It is irreversible and acts on free tubulin as well as microtubules.||ZM 449829 - Potent transglutaminase inhibitor|
|Microtubule associated proteins (MAPs)||Microtubule lattice||Associates with microtubules and promote assembly and stabilization.|
|Plus-end tracking proteins (+TIPs)||+TIP recruitment results in microtubule stabilization, continued growth, and protrusion of the plus ends (migration).|
|Tubulin-binding agents||Depolymerizing agents||Inhibits microtubule polymerization, block mitosis and induces cell death. Most agents bind the vinca or colchicine domain.|
|Stabilizing agents||Disrupts microtubules by promoting polymerization. This also inhibits mitosis and induces cell death.|
Acetylation detection: Immunocytochemistry of formaldehyde-fixed HeLa cells stained with anti-alpha tubulin (acetyl K40) antibody [EPR16772] (ab179484). Positive control - Cell treatment with trichostatin A (ab120850), a potent deacetylase inhibitor. Nuclear DNA was labeled with DAPI (shown in blue). Click here to view the full datasheet for ab179484.
|Motility||Kinesin||Aided by ATP, they shuttle vesicular cargo away from the MTOC towards the plus-end of the microtubule.|
|Dynein||Aided by ATP, they shuttle vesicular cargo towards the minus end of microtubules.||Dynein mouse monoclonal antibody|
|Myosin||Mainly associates with actin, but also interacts with tubulin for cargo exchange.|
|Mitosis||Kinetochore||Interacts with microtubules to maintain the integrity of chromatid spindles.|
|Cell migration||Actin||Crosstalk between actin and microtubules promotes symmetry break to polarize cells for division, shape changes, and migration.|
HeLa cells were stained with mouse anti-tubulin followed with a fluorescent red Goat Anti-Mouse IgG, actin filaments were stained with CytoPainter Phalloidin-iFluor 488 Reagent (ab176753), and nuclei were stained with Hoechst 33342.