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Anisomycin

別名：Flagecidin, Wuningmeisu C

Anisomycin (Flagecidin, Wuningmeisu C) is a bacterial antibiotic isolated from Streptomyces griseolus, which inhibits protein synthesis, and also act as a JNK activator. Anisomycin upregulates autophagy and increases apoptosis.

Anisomycin化学構造

CAS No. 22862-76-6

サイズ 価格(税別) 在庫状況
10mM (1mL in DMSO) JPY 29500 国内在庫あり
JPY 22000 国内在庫あり
JPY 44500 国内在庫あり
JPY 85500 国内在庫あり
JPY 220500 国内在庫なし(納期7~10日)

代表番号: 045-509-1970|電子メール：[email protected]
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Anisomycin関連製品

シグナル伝達経路

JNK Signaling Pathways

JNKシグナル伝達経路

JNK阻害剤の選択性比較

Cell Data

Cell Lines Assay Type Concentration Incubation Time 活性情報 PMID
HEK293 Function assay 100 uM 15 mins Increase of JNK phosphorylation in U50488 treated untransfected HEK293 cells at 100 uM after 15 mins 17702750
HEK293 Function assay 50 uM 15 mins Increase of U50488-induced JNK phosphorylation in untransfected HEK293 cells at 50 uM after 15 mins 17702750
RAW264.7 Function assay 5 uM 30 mins Activation of p38MAPK in mouse RAW264.7 cells assessed as phosphorylation at Thr180/Tyr182 at 5 uM after 30 mins by Western blotting analysis 23294286
Sf9 Function assay 10 uM 6 to 24 hr Increase of ATP level in Spodoptera frugiperda (fall armyworm) Sf9 cells at 10 uM after 6 to 24 hr by luminescent cell viability assay ChEMBL
Sf9 Cytotoxicity assay 10 uM 72 hr Cytotoxicity against Spodoptera frugiperda (fall armyworm) Sf9 cells at 10 uM after 72 hr by trypan blue dye exclusion test ChEMBL
HeLa Function assay 10 uM Inhibition of translation in human HeLa cells at 10 uM by 35S-methionine metabolic labeling study 15165136
VERO-E6 Function assay 48 hrs Determination of IC50 values for inhibition of SARS-CoV-2 induced cytotoxicity of VERO-E6 cells after 48 hours exposure to 0.01 MOI SARS CoV-2 virus by high content imaging, IC50=0.09μM. ChEMBL
VERO-E6 Function assay 48 hrs Toxicity CC50 against VERO-E6 cells determined at 48 hours by high content imaging (same conditions as 2_LEY without exposure to 0.01 MOI SARS CoV-2 virus), CC50=0.1μM. ChEMBL
HEK293 Function assay Inhibitory concentration required to produce cytotoxicity against HEK293 cells, IC50=0.02μM. 16005213
Vero E6 Function assay CC50 determination at MOI 0.004 using CellTiter- Glo (CTG) assay, performed 3 days post-infection in Vero E6 cells, CC50<0.39μM. ChEMBL
Vero E6 Function assay CC50 determination at MOI 0.01 using CellTiter- Glo (CTG) assay, performed 3 days post-infection in Vero E6 cells, CC50<0.39μM. ChEMBL
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生物活性

製品説明 Anisomycin (Flagecidin, Wuningmeisu C) is a bacterial antibiotic isolated from Streptomyces griseolus, which inhibits protein synthesis, and also act as a JNK activator. Anisomycin upregulates autophagy and increases apoptosis.
Targets
JNK [1]
In Vitro
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.[2] 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.[3] Anisomycin inhibits EAC cell proliferation in concentration-dependent manner.[4]
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 (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.
細胞実験 細胞株 Ehrlich ascites carcinoma (EAC) cells
濃度 500 ng/mL
反応時間 48 h
実験の流れ 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.
実験結果図 Methods Biomarkers 結果図 PMID
Western blot p-Keratin 20 / Keratin 20 / p-Keratin 18 / Keratin 18 / p-Keratin 8 / Keratin 8 / p-MK2 / p-hHSPB1 / hHSPB1 PP2A / PP2C p-ERK / ERK / p-JNK / JNK / p-p38 / ATF3 20724476
Immunofluorescence p-Keratin 8 / p-Keratin 18 / p-Keratin 20 20724476
Growth inhibition assay Cell viability 22684030
In Vivo
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.[4]
動物実験 動物モデル Male BALB/c mice
投与量 5 mg/kg
投与経路 peritumorally

化学情報

分子量 265.3 化学式

C14H19NO4
CAS No. 22862-76-6 SDF Download Anisomycin SDFをダウンロードする
Smiles CC(=O)OC1C(CNC1CC2=CC=C(C=C2)OC)O
保管

In vitro
Batch:

DMSO : 53 mg/mL ( (199.77 mM)； 吸湿したDMSOは溶解度を減少させます。新しいDMSOをご使用ください。)

Ethanol : 16 mg/mL

Water : Insoluble

モル濃度計算器

in vivo
Batch:

Add solvents to the product individually and in order.

投与溶液組成計算機

実験計算

モル濃度計算器

質量 濃度 体積 分子量

投与溶液組成計算機(クリア溶液)

ステップ１：実験データを入力してください。（実験操作によるロスを考慮し、動物数を1匹分多くして計算・調製することを推奨します）

mg/kg g μL 匹

ステップ２：投与溶媒の組成を入力してください。（ロット毎に適した溶解組成が異なる場合があります。詳細については弊社までお問い合わせください）

% DMSO % % Tween 80 % ddH2O
%DMSO %

計算結果：

投与溶媒濃度： mg/ml;

DMSOストック溶液調製方法： mg 試薬を μL DMSOに溶解する（濃度 mg/mL, 注：濃度が当該ロットのDMSO溶解度を超える場合はご連絡ください。 )

投与溶媒調製方法：Take μL DMSOストック溶液に μL PEG300，を加え、完全溶解後μL Tween 80，を加えて完全溶解させた後 μL ddH2O，を加え完全に溶解させます。

投与溶媒調製方法：μL DMSOストック溶液に μL Corn oil，を加え、完全溶解。

注意：１.ストック溶液に沈殿、混濁などがないことをご確認ください；
２.順番通りに溶剤を加えてください。次のステップに進む前に溶液に沈殿、混濁などがないことを確認してから加えてください。ボルテックス、ソニケーション、水浴加熱など物理的な方法で溶解を早めることは可能です。

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