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Biological Description

Description: Anisomycin is an antibiotic isolated from various Streptomyces species. It interferes with protein and DNA synthesis by inhibiting peptidyl transferase or the 80S ribosome system.

In vitro: 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.

In vivo: Peritumoral 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.

Kinase Assay: JNK phosphorylation: 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 (?nal concentration 1% v/v). Puromycin is added (?nal 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 ?uorescence signal for anti-phospho-JNK antibody is background corrected and normalized for loading.

Cell Research: For the assay, 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.(Only for Reference)

Chemical Properties

Molecular Weight: 265.3

Formula: C14H19NO4

CAS No.: 22862-76-6

Storage & Solubility Information

Storage

Powder: -20°C for 3 years

In solvent: -80°C for 2 years

Solubility Information

Ethanol: 13.3 mg/mL (50 mM)

DMSO: 26.5 mg/mL (100 mM)

( < 1 mg/ml refers to the product slightly soluble or insoluble )

Catalog #: T6758

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References and Literature

1. Iordanov MS, et al. Mol Cell Biol. 1997, 17(6), 3373-3381.

2. Monaqhan D, et al. Biochem Biophys Res Commun. 2014, 443(2), 761-767.

3. Li JY, et al. Acta Pharmacol Sin. 2012, 33(7), 935-940.

4. You P, et al. Oncol Rep. 2013, 29(6), 2227-2236.

5. Chen L, Zhou X, Kong X, et al. The Prognostic Significance of Anisomycin-Activated Phospho-c-Jun NH2-Terminal Kinase (p-JNK) in Predicting Breast Cancer Patients’ Survival Time[J]. Frontiers in Cell and Developmental Biology. 2021, 9: 470.

6. Liu Y J, Chang Y J, Kuo Y T, et al. Targeting β-tubulin/CCT-β complex induces apoptosis and suppresses migration and invasion of highly metastatic lung adenocarcinoma[J]. Carcinogenesis. 2020, 41(5): 699-710.