Cyclopamine

Manufactured by R&D Systems

Sourced in United States

Cyclopamine is a naturally occurring alkaloid compound that has been studied for its effects on cellular signaling pathways. It is commonly used as a research tool in cell and molecular biology studies.

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Lab products found in correlation

Percoll solution, by Merck Group (3 mentions) Poly d lysine, by Merck Group (3 mentions) Matrigel, by BD (3 mentions) Poly dl ornithine, by Merck Group (3 mentions) Hepg2, by American Type Culture Collection (1 mentions) Capan 1, by American Type Culture Collection (1 mentions) L3.6pl, by American Type Culture Collection (1 mentions) Nih3t3 e1 fibroblasts, by American Type Culture Collection (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Skbr3, by American Type Culture Collection (1 mentions) Mda mb 231, by American Type Culture Collection (1 mentions) C3h10t1 2, by American Type Culture Collection (1 mentions) Gant61, by R&D Systems (1 mentions) Cytochalasin d, by MP Biomedicals (1 mentions)

5 protocols using cyclopamine

Generation and Characterization of Medulloblastoma Cells

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CGNP cultures were generated as described previously (33 (link)). Cells were plated on poly-DL-ornithine (Sigma) precoated plates or precoated glass coverslips. Where indicated, Shh was used at a concentration of 3 μg/mL. Cyclopamine (R&D Systems) was used at 1 μg/mL.
Medulloblastoma cells (MBC) were harvested from SmoA1 mouse medulloblastomas. Briefly, tumors were disassociated and cells were incubated in Papain/DNAse solution solution for 30 minutes, and then passed through a cell strainer. Cells were subsequently separated on a density step gradient of 35% and 60% Percoll solution (Sigma). Purified MBCs were enriched by pre-plating on uncoated tissue culture dishes to remove adherent fibroblasts and glial cells. Non-adherent cells were plated on tissue culture dishes or glass coverslips pre-coated with poly D-lysine (Sigma) and Matrigel (BD Biosciences), infected with adenoviruses and cultured for 48 hours before western blotting.

Malhotra A., Dey A., Prasad N, & Kenney A.M. (2015). Sonic Hedgehog signaling drives mitochondrial fragmentation by suppressing mitofusins in cerebellar granule neuron precursors and medulloblastoma. Molecular cancer research : MCR, 14(1), 114-124.

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Cell Culture Protocols for Various Cell Lines

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C3H10T1/2 mouse embryonic fibroblasts and NIH3T3-E1 fibroblasts were obtained from ATCC (Manassas, VA) and cultured as previously described [Dwyer et al., 2007 (link); Richardson et al., 2007 (link)]. CAPAN-1, L3.6pl, and E3LZ10.7 human pancreatic cancer cells were obtained from ATCC and cultured in DMEM containing 10% heat-inactivated fetal bovine serum (FBS) (Hyclone Laboratories, Logan, UT). The Human hepatoma cell line HepG2 was obtained from ATCC and cultured in DMEM containing 10% FBS. Mouse SUFU null (−/−) MEFs were provided by Dr. Philip Beachy at Stanford University and cultured in DMEM containing 10 FBS as previously described. LXRαβ null (−/−) MEFs were provided by Dr. Peter Tontonoz at UCLA and cultured in DMEM containing 10% FBS. GH3 cells were obtained from ATCC and cultured in Hamʹs F10 containing 2 mM Glutamine, 15% Horse Serum and 2.5 FBS. TO901317 LXR agonist was purchased from Cayman Chemicals (Ann Arbor, MI). Recombinant human Shh N-terminal peptide and cyclopamine were obtained from R&D Systems, Inc. (Minneapolis, MN), and Shh-neutralizing antibody 5E1 was obtained from Developmental Studies Hybridoma Bank (Iowa City, IA).

Wang F., Stappenbeck F., Matsui W, & Parhami F. (2016). Inhibition of Pancreatic Cancer Cell-Induced Paracrine Hedgehog Signaling by Liver X Receptor Agonists and Oxy16, a Naturally Occurring Oxysterol. Journal of cellular biochemistry, 118(3), 499-509.

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Culturing Breast Cancer Cell Lines

Cited 1 time

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The SUM149, SUM159 and SUM190 cell lines were obtained from Asterand Inc (Detroit, MI) and cultured per manufacturer’s instructions and as described previously [35 (link)]. MDA-MB-231, SKBR3 cells and C3H10T1/2 cell lines were from American Type Culture Collection (ATCC) (Manassas, VA) and cultured per their instructions. Human mammary epithelial cells (HMEC) were obtained from Lonza Group Ltd. (Allendale, NJ) and cultured per manufacturer’s instructions using their Mammary Epithelial Growth Medium Kit (CC-3150). GANT58 (Cat # 3889), GANT61 (Cat # 3191), JK184 (Cat # 3341) and cyclopamine were obtained from Tocris/R&D Systems (Minneapolis, MN). GANT61 in gram quantities was from DC Chemicals (Shanghai, China). HPI-1/2/3/4 were a kind gift from Dr. James K. Chen, Stanford School of Medicine. HPI-1/3/4 were also purchased from Sigma Aldrich (St. Louis, MO). Cytochalasin D was from MP Biomedicals, LLC (Solon, OH). Hoechst-33342 and YOYO-1 dyes were from Thermo Fisher Scientific

Oladapo H.O., Tarpley M., Sauer S.J., Addo K.A., Ingram S.M., Strepay D., Ehe B.K., Chdid L., Trinkler M., Roques J.R., Darr D.B., Fleming J.M., Devi G.R, & Williams K.P. (2017). Pharmacological targeting of GLI1 inhibits proliferation, tumor emboli formation and in vivo tumor growth of inflammatory breast cancer cells. Cancer letters, 411, 136-149.

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Medulloblastoma Cell Culture and Analysis

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CGNP cultures were generated as described previously (Kenney and Rowitch 2000). Cells were plated on poly-DL-ornithine (Sigma, St Louis, MO, USA) precoated plates or precoated glass coverslips. Where indicated, Shh was used at a concentration of 3 μg/mL. Cyclopamine (R&D Systems, Minneapolis, MN, USA) was used at 1 μg/mL. Medulloblastoma cells (MBC) were harvested from SmoA1 mouse medulloblastomas. Briefly, tumors were disassociated and cells were incubated in Trypsin/EDTA solution for 30 minutes, then passed through a cell strainer. Cells were subsequently separated on a density step gradient of 35% and 65% Percoll solution (Sigma, St Louis, MO, USA). Purified MBCs were enriched by pre-plating on uncoated tissue culture dishes to remove adherent fibroblasts and glial cells. Non-adherent cells were plated on tissue culture dishes pre-coated with poly D-lysine (Sigma, St Louis, MO, USA) and Matrigel (BD Biosciences, San Jose, CA), infected with adenoviruses/lentiviruses 4 hours later, and cultured for 72 more hours before collection for further analysis. Mouse medulloblastoma derived PZp53Med cell line (38 (link)) was used in the ChIP experiments (below) and the cells were mycoplasma free

Dey A., Robitaille M., Remke M., Maier C., Malhotra A., Gregorieff A., Wrana J.L., Taylor M.D., Angers S, & Kenney A.M. (2016). YB-1 is elevated in medulloblastoma and drives proliferation in Sonic hedgehog - dependent cerebellar granule neuron progenitor cells and medulloblastoma cells. Oncogene, 35(32), 4256-4268.

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Medulloblastoma Cell Culture and Analysis

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