Colchicine

Manufactured by Enzo Life Sciences

Sourced in United States

Colchicine is a laboratory reagent used for various applications in cell and molecular biology research. It is a natural alkaloid compound extracted from plants of the Colchicum genus. Colchicine is commonly used to inhibit cell division and disrupt microtubule formation, which has applications in areas such as cytogenetics and cell cycle analysis.

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3 protocols using colchicine

Pharmacological Modulation of Cytoskeletal Networks

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Specific pharmacological agents were first used to inhibit the polymerization of microtubules25 (link) (Colchicine, Enzo Life Sciences, Zandhoven, Belgium), to disorganize neurofilaments26 (link) (ß-ß'-iminodipropionitrile, IDPN, Sigma Aldrich, St Louis, MO) and actin filaments27 (latrunculin-A, Enzo Life Sciences, Zandhoven, Belgium). Colchicine was applied at 1 μM for 20 min, IDPN at 5% w/v for 20 min. and latrunculin at 10 μM for 30 min. In addition, we used Blebbistatin (Enzo Life Sciences, Zandhoven, Belgium) at 75 μM during 20 min. to inhibit myosin II contractility61 (link), monastrol (Sigma Aldrich, St Louis, MO) at 100 μM during 30 min. to inhibit the ATPase activity of kinesin 562 (link), erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA, Sigma Aldrich, St Louis, MO) at 100 μM during 20 min. to inhibit the ATPase activity of the cytoplasmic dynein63 (link) and 2,5-hexanedione (Sigma Aldrich, St Louis, MO) at 10 mM during 30 min. to increase the cross-linking of neurofilaments64 (link). Control experiments were performed with DMSO up to 0.5% during 30 min. and additional experiments demonstrated that the effects of all of these pharmacological agents were sustained for the entire experimental period.

Grevesse T., Dabiri B.E., Parker K.K, & Gabriele S. (2015). Opposite rheological properties of neuronal microcompartments predict axonal vulnerability in brain injury. Scientific Reports, 5, 9475.

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Synthesis and Evaluation of SKI-178 Compound

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SKI-178
(N′-[(1E)-1-(3,4-dimethoxyphenyl)ethylidene]-3-(4-methoxxyphenyl)-1H-pyrazole-5-carbohydrazide)
was synthesized as described previously.^{24 (link)} Reagents were purchased as follows:vincristine (Thermo
Fisher Scientific, Waltham, MA, USA), Colchicine (Enzo Life Sciences,
Farmingdale, NY, USA), Paclitaxel (Sigma-Aldrich, St. Louis, MO, USA), MG132
(Sigma-Aldrich, St. Louis, MO, USA) and PF-543 (Selleck Chemicals, Houston, TX,
USA), antibodies against SphK1, pBcl-2(Ser70), Cleaved caspase-7, and Tubulin
(Cell Signaling Technology, Danvers, MA, USA), SphK1 (pSer225) (ECM Biosciences,
Versailles, KY), and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA).
β-hydroxy-propyl-cyclodextrin was from Acros (NJ, USA). 1,2-Propanediol,
Tween-20, and dextrose were from Sigma Aldrich (MO, USA).

Hengst J.A., Dick T.E., Sharma A., Doi K., Hegde S., Tan S.F., Geffert L.M., Fox T.E., Sharma A.K., Desai D., Amin S., Kester M., Loughran T.P., Paulson R.F., Claxton D.F., Wang H.G, & Yun J.K. (2017). SKI-178: A Multitargeted Inhibitor of Sphingosine Kinase and Microtubule Dynamics Demonstrating Therapeutic Efficacy in Acute Myeloid Leukemia Models. Cancer translational medicine, 3(4), 109-121.

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Lipid-based Nanoparticle Transfection Protocol

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The phospholipids, dioleoyl-3-trimethylammonium-propane chloride (DOTAP), dioleoyl-snglycero-3-phosphoethanolamine (DOPE), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) were purchased from Avanti Polar Lipids (Birmingham, AL, USA). The phospholipid DSPE–PEG₂₀₀₀–NHS was purchased from Biochempeg Scientific Inc (Watertown, MA, USA). Holo-transferrin bovine, cholesterol, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), Ethylenediaminetetraacetic acid (EDTA), 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), Hoechst 33342, Ethidium bromide (EtBr), Sodium azide, Amiloride and Triton™ X-100 were obtained from Sigma–Aldrich (St. Louis, MO, USA). Chlorpromazine and Colchicine were obtained from Enzo Life Sciences (Farmingdale, NY, USA). Chitosan (MW 30 kDa) was purchased from Glentham Life Sciences (Corsham, UK). Plasmid DNA encoding β-galactosidase (gWiz-βGal) and plasmid DNA encoding Green Fluorescent Protein (gWiz-GFP) were purchased from Aldevron LLC (Fargo, ND, USA). Dulbecco’s Modified Eagle Medium (DMEM), and phosphate buffered saline (PBS) were purchased from Corning Incorporated (Corning, NY, USA). Fetal bovine serum (FBS) was purchased from JR Scientific Inc. (Woodland, CA, USA). β-galactosidase enzyme assay kit with reporter lysis buffer was supplied by Promega (Madison, WI, USA).

dos Santos Rodrigues B., Banerjee A., Kanekiyo T, & Singh J. (2019). Functionalized Liposomal Nanoparticles for Efficient Gene Delivery System to Neuronal Cell Transfection. International journal of pharmaceutics, 566, 717-730.

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