Ciliobrevin a

Manufactured by Bio-Techne

Sourced in United States

Ciliobrevin A is a chemical compound that functions as a selective inhibitor of cytoplasmic dynein, a motor protein responsible for the movement of cellular structures, including cilia and flagella. This compound can be used in research applications to study the role of cytoplasmic dynein in various cellular processes.

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Lab products found in correlation

Rhodamine phalloidin, by Thermo Fisher Scientific (3 mentions) Bi2536, by Selleck Chemicals (1 mentions) Tubacin, by Enzo Life Sciences (1 mentions) Nocodazole, by Merck Group (1 mentions) Tubastatin a, by Merck Group (1 mentions) Colchicine, by Merck Group (1 mentions) Forskolin, by Bio-Techne (1 mentions) Cyclopamine, by Cayman Chemical (1 mentions) Palbociclib, by Cayman Chemical (1 mentions) Torin1, by Bio-Techne (1 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions) Prolonged diamond dapi, by Thermo Fisher Scientific (1 mentions) Mln4924, by MedChemExpress (1 mentions) 4 hydroxytamoxifen, by Merck Group (1 mentions)

4 protocols using ciliobrevin a

Embryo Asymmetry Disruption Assay

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For chemical treatment, embryos were treated with final 2 μg/mL PTX for Gαi inhibition^{52 (link)} (P2980, Sigma), 25 μg/mL SDS to disrupt asymmetry, 100 μM Ciliobrevin A to inhibit cytoplasmic AAA + ATPase dynein-dependent microtubule gliding and ATPase activity^{53 (link)} (TOCRIS bioscience), or 10 μM BI2536 to block PLK1 activity (Selleckchem, USA) during the 8–16 cell stage for 10 min before fixation for immunofluorescence. For phalloidin staining, embryos were fixed with 3.7% paraformaldehyde for 15 min, followed by PBS wash three times with 0.1% triton, incubated with 10 unit/mL of Rhodamine-Phalloidin (Thermo Fisher Scientific) for 20 min, washed with PBS three times all at room temperature prior to imaging. The details of the reagents are also listed in Supplementary Table 8.

Poon J., Fries A., Wessel G.M, & Yajima M. (2019). Evolutionary modification of AGS protein contributes to formation of micromeres in sea urchins. Nature Communications, 10, 3779.

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Inhibition of NLRP3 Inflammasome Activation

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We pre-treated LPS-primed iBMDMs for 1 hour with HDAC6 inhibitors 10 μM Tubastatin A (Sigma-Aldrich, Cat. no: SML0044), 30 μM Rocilinostat (Selleckchem, Cat. no: S8001), or 20 μM, or 5–40 μM Tubacin (Enzo Life Sciences, Cat. no: BML-GR362–0500), 10 μM microtubule polymerization inhibitors Colchicine (Sigma-Aldrich, Cat. no: C9754–100MG) or 10 μM Nocodazole (Sigma-Aldrich, Cat. no: M1404–2MG), or with 0.1–20 μM NLRP3 inhibitor MCC950 (CP-456773) (Cayman Chemicals Cat. no: 210826–40-7 and Sigma-Aldrich, Cat. no: PZ0280). Cytoplasmic dynein-dependent microtubule transport was inhibited using 25 μM Ciliobrevin A (TOCRIS bioscience, Cat. no. 4529). One-hour post drug pre-treatment, activation was carried out either with nigericin (30 min), dsDNA (6 hours), or TcdB (1 hour). iBMDMs were pretreated with 5 mM of 3-MA (Invivogen, Cat. no: tlrl-3ma) for 6 hours to block autophagy.

Magupalli V.G., Negro R., Tian Y., Hauenstein A.V., Caprio G.D., Skillern W., Deng Q., Orning P., Alam H.B., Maliga Z., Sharif H., Hu J.J., Evavold C.L., Kagan J.C., Schmidt F.I., Fitzgerald K.A., Kirchhausen T., Li Y, & Wu H. (2020). HDAC6 mediates an aggresome-like mechanism for NLRP3 and pyrin inflammasome activation. Science (New York, N.Y.), 369(6510), eaas8995.

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Modulation of Hedgehog Signaling

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The following small molecules were used and dissolved in DMSO, ethanol, or water according to product data sheets as 1,000× stocks: cyclopamine (Cayman Chemical), SAG (Cayman chemical), Pr-8-Bromo cAMPs (sodium salt; Cayman Chemical), purmorphamine (Stemgent), GSA-10 (Tocris), palbociclib (Cayman Chemical), Torin 1, Ciliobrevin A (Tocris), and forskolin (Tocris). Small molecules were added to the cells 2 h before Hh induction and remained in the Shh CM throughout the experiment unless otherwise indicated.

Akhshi T, & Trimble W.S. (2020). A non-canonical Hedgehog pathway initiates ciliogenesis and autophagy. The Journal of Cell Biology, 220(1), e202004179.

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Pharmacological Modulation of Ciliogenesis

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Ciliobrevin A was purchased from Tocris (#4529) and MLN4924 (Pevonedistat, # HY-70062) was purchased from MedChemExpress (MCE). Paraformaldehyde (#J19943-K2) and Prolonged Diamond DAPI (#P36966) were purchased from Thermofisher. Finally, 4-hydroxytamoxifen was purchased from Sigma (#H6278) and SAG was purchased from Abcam (#ab142160). The working concentration of: a) Ciliobrevin A is 50µM, b) SAG is 1µg/ml, c) 4-hydroxytamoxifen for the ex vivo experiments is 2µg/ml and D) MLN4924 is 0.25 or 0.5μM.
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.

Petsouki E., Gerakopoulos V., Szeto N., Chang W., Humphrey M.B., & Tsiokas L. (2020). FBW7 couples structural integrity with functional output of primary cilia.

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