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    Anisomycin (NSC-76712, AI 3-50846, Flagecidin, Wuningmeisu C)
    Anisomycin (NSC-76712, AI 3-50846, Flagecidin, Wuningmeisu C)

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    This product is for research use only, not for human use. We do not sell to patients.
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    InvivoChem Cat #: V0475
    CAS #: 22862-76-6Purity ≥98%

    Description: Anisomycin (Wuningmeisu C, NSC76712, AI3-50846, Flagecidin), a naturally occuring bacterial antibiotic isolated from Streptomyces griseolus, is a novel, potent and specific activator (agonist) of JNK (c-Jun N-terminal Kinase) with potential antineoplastic activity. It is also a potent and reversible inhibitor of protein synthesis in eukaryotic organisms that acts by binding and inhibiting peptidyl transferase activity of 60S ribosomal subunit.

    References: Mol Cell Biol. 1997 Jun;17(6):3373-81; Biochem Biophys Res Commun. 2014 Jan 10;443(2):761-7.

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    Molecular Weight (MW)265.3
    CAS No. 22862-76-6
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 41 mg/mL (154.5 mM)          
    Water:<1 mg/mL 
    Ethanol: 17 mg/mL warmed (64.1 mM) 
    Solubility (In vivo) 2% DMSO+corn oil: 5mg/mL 

    Wuningmeisu C; NSC 76712; AI3 50846; NSC-76712; AI-350846; NSC76712; AI350846; Flagecidin; 

    Chemical Name: (2R,3S,4S)-4-hydroxy-2-(4-methoxybenzyl)pyrrolidin-3-yl acetate


    InChi Code: InChI=1S/C14H19NO4/c1-9(16)19-14-12(15-8-13(14)17)7-10-3-5-11(18-2)6-4-10/h3-6,12-15,17H,7-8H2,1-2H3/t12-,13+,14+/m1/s1

    SMILES Code: O[[email protected]@H]1[[email protected]@H](OC(C)=O)[[email protected]@H](CC2=CC=C(OC)C=C2)NC1 

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    In Vitro

    In vitro activity: Anisomycin (3 μM) decreases protein synthesis in MDA16 and MDA-MB-468 cells, and reduces colony formation by MDA-MB-468 cells. Anisomycin causes an increase in the number of apoptotic cells in MDA-MB-468 cultures, but not in MDA16 cultures. Anisomycin actives JNK phosphorylation in MDA-MB-468 cells. In U251 and U87 cells, anisomycin (0.01-8 μM) inhibits the cell growth in time- and concentration-dependent manners with the IC50 (48 h) values of 0.233 and 0.192 μmol/L, respectively. Anisomycin (4 μM) causes 21.5% and 25.3% of apoptosis proportion in U251 and U87 cells, respectively, and activates p38 MAPK and JNK, while inactivated ERK1/2. Anisomycin (4 μM) reduces the level of PP2A/C subunit in a time-dependent manner in U251 and U87 cells. Anisomycin inhibits EAC cell proliferation in concentration-dependent manner.

    Kinase Assay: Cells (500,000 cells/well )are seeded in 6-well plates and incubated overnight. Cells are then incubated for 1 h with test compounds or DMSO as vehicle control (final concentration 1% v/v). Puromycin is added (final concentration of 18 μM) and cells incubated for a further 10 min to label nascent polypeptide chains. Background labelling is determined by incubating cells without puromycin. Cells are then washed in HBSS, harvested by scraping and centrifuged (300 g, 5 min). Cells are resuspended in 0.5 mL 50 mM DTT containing phosphatase inhibitors and incubated at 95℃ for 10 min. Samples are then snap frozen in liquid nitrogen and stored at -20℃ until blotted. Samples (20–30 μg protein/sample) are blotted onto a PVDF membrane. The membrane is blocked and incubated with anti-phospho-Thr183/Tyr185-JNK antibody overnight at 4℃. Secondary antibodies are used to label the primary antibody and detected using an infrared scanner. The intensity of the fluorescence signal for anti-phospho-JNK antibody is background corrected and normalized for loading.

    Cell Assay: For the assay, Ehrlich ascites carcinoma (EAC) cells are plated in 96-well plates at a density of 10,000 cells/well/200 µL of medium. The cells are treated with the different concentrations of anisomycin for 48 h. Adriamycin (500 ng/mL) is used as a positive control. 0.5 mg/mL of MTT is added to each well. 4 h later, the formazan product of MTT reduction is dissolved in DMSO, and absorbance is measured at 570 nm using a Model 680 microplate reader.

    In VivoPeritumoral administration of anisomycin (5 mg/kg) significantly suppresses Ehrlich ascites carcinoma (EAC) growth resulting in the survival of approximately 60% of the mice 90 days after EAC inoculation.
    Animal modelMale BALB/c mice
    Formulation & DosageDissolved in PBS; 5 mg/kg; Administered peritumorally

    Mol Cell Biol. 1997 Jun;17(6):3373-81; Oncol Rep. 2013 Jun;29(6):2227-36. 

    These protocols are for reference only. InvivoChem does not independently validate these methods.


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