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5mg |
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25mg |
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50mg |
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Purity: ≥98%
Cyclopamine (formerly HSDB3505; HSDB-3505; 11-deoxojervine), a naturally occuring steroid alkaloid found in Veratrum plant, is reported to be a potent and specific inhibitor of Hedgehog (Hh) signaling pathway with both anticancer and teratogenic activities. In TM3Hh12 cells, it inhibits Smoothened (Smo) at an IC50 of 46 nM.
Targets |
Human Endogenous Metabolite
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ln Vitro |
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ln Vivo |
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Enzyme Assay |
Using Luciferase as a readout, the Gli-Luc assay measures the transcriptional modulation of Gli, the final stage of the Hh signaling pathway. After serial dilution in DMSO, cyclopamine is ready for assay and added to assay plates that are empty. After being resuspended in F12 Ham's/DMEM (1:1) containing 5% FBS and 15 mM Hepes pH 7.3, TM3Hh12 cells (TM3 cells with the Hh-responsive reporter gene construct pTA-8xGli-Luc) are added to assay plates and incubated with Cyclopamine for about 30 minutes at 37°C in 5% CO2. After that, assay plates are filled with 1 nM Hh-Ag 1.5 and allowed to incubate at 37 °C with 5% CO2. Following 48 hours, the assay plates are refilled with either Bright-Glo or MTS reagent, and the absorbance or luminescence at 492 nm is measured. The logistic curve's inflection point, or IC50 value, is found by non-linearly regressing the Gli-driven luciferase luminescence or absorbance signal from the MTS assay against log10 (cyclopamine concentration) using the R statistical software package.
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Cell Assay |
In 96-well plates, cells are exposed to cyclopamine. Soluble tetrazolium salt, or MTS, assay is used to measure cell viability. Using the CellTiter96 colorimetric assay, optical density measurements at 490 nm (OD490) at 2 and 4 days determine the viable cell mass. The formula for calculating relative growth is OD (day 4)⋣OD (day 2)/OD (day 2).
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Animal Protocol |
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References |
Molecular Formula |
C27H41NO2
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Molecular Weight |
411.62
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Exact Mass |
411.31
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Elemental Analysis |
C, 78.78; H, 10.04; N, 3.40; O, 7.77
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CAS # |
4449-51-8
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Related CAS # |
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Appearance |
Solid powder
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SMILES |
C[C@H]1C[C@@H]2[C@H]([C@H]([C@]3(O2)CC[C@H]4[C@@H]5CC=C6C[C@H](CC[C@@]6([C@H]5CC4=C3C)C)O)C)NC1
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InChi Key |
QASFUMOKHFSJGL-LAFRSMQTSA-N
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InChi Code |
InChI=1S/C27H41NO2/c1-15-11-24-25(28-14-15)17(3)27(30-24)10-8-20-21-6-5-18-12-19(29)7-9-26(18,4)23(21)13-22(20)16(27)2/h5,15,17,19-21,23-25,28-29H,6-14H2,1-4H3/t15-,17+,19-,20-,21-,23-,24+,25-,26-,27-/m0/s1
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Chemical Name |
(3S,3'R,3'aS,6'S,6aS,6bS,7'aR,9R,11aS,11bR)-3',6',10,11b-tetramethylspiro[2,3,4,6,6a,6b,7,8,11,11a-decahydro-1H-benzo[a]fluorene-9,2'-3a,4,5,6,7,7a-hexahydro-3H-furo[3,2-b]pyridine]-3-ol
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Synonyms |
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HS Tariff Code |
2934.99.9001
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Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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Solubility (In Vitro) |
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Solubility (In Vivo) |
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Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 2.4294 mL | 12.1471 mL | 24.2943 mL | |
5 mM | 0.4859 mL | 2.4294 mL | 4.8589 mL | |
10 mM | 0.2429 mL | 1.2147 mL | 2.4294 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration: mg/mL;
Method for preparing DMSO stock solution: mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation::Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
A, viable cell masses, determined using MTT assays, were reduced by nearly 100% to 0% by Hh inhibition with 6 μmol/L cyclopamine for 4 d as compared with solvent-treated cells in vitro. Cancer Res . 2007 Mar 1;67(5):2187-96. td> |
A, using the Guava Multicaspase assay, an increase of cells undergoing apoptosis was detected on treatment with cyclopamine. Cancer Res . 2007 Mar 1;67(5):2187-96. td> |
Effects of cyclopamine treatment on pancreatic adenocarcinoma cells. Nature . 2003 Oct 23;425(6960):851-6. td> |
Cyclopamine treatment blocks tumour formation of human pancreatic adenocarcinoma cells after transplantation into nude mice. Nature . 2003 Oct 23;425(6960):851-6. td> |