Name: Anisomycin (Flagecidin)
Brand: Selleck Chemicals
SKU: S7409
Availability: InStock
Rating: 5 (85 reviews)

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Quality Control

Batch: Purity: 99.93%

99.93

Related Targets	ERK p38 MAPK Raf MEK Ras KRas S6 Kinase MAP4K TAK1 Mixed Lineage Kinase
Other JNK Inhibitors	CC-90001 SP600125 JNK-IN-8 JNK Inhibitor IX Tanzisertib Hydrochloride (CC-930) JNK Inhibitor VIII Bentamapimod (AS602801) Polyphyllin I DTP3 BI-78D3

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
HEK293	Function assay			Inhibitory concentration required to produce cytotoxicity against HEK293 cells, IC50=0.02μM.	16005213
HeLa	Function assay	10 uM		Inhibition of translation in human HeLa cells at 10 uM by 35S-methionine metabolic labeling study	15165136
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
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
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
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 53 mg/mL (199.77 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 16 mg/mL

Water : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	2.650mg/ml (9.99mM)	Taking the 1 mL working solution as an example, add 50 μL of 53 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.850mg/ml (3.20mM)	Taking the 1 mL working solution as an example, add 50 μL of 17 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg

Average weight of animals: g

Dosing volume per animal: μL

Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO

%

% Tween 80

% ddH₂O

% DMSO

+

%

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Chemical Information, Storage & Stability

Molecular Weight	265.3	Formula	C₁₄H₁₉NO₄	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	22862-76-6	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent

Mechanism of Action

Targets/IC₅₀/Ki	JNK
In vitro	Anisomycin (3 μM) decreases protein synthesis in MDA16 and MDA-MB-468 cells, and reduces colony formation by MDA-MB-468 cells. This compound causes an increase in the number of apoptotic cells in MDA-MB-468 cultures, but not in MDA16 cultures. It actives JNK phosphorylation in MDA-MB-468 cells. In U251 and U87 cells, this chemical (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. It (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. This compound (4 μM) reduces the level of PP2A/C subunit in a time-dependent manner in U251 and U87 cells. It inhibits EAC cell proliferation in concentration-dependent manner.
Kinase Assay	JNK phosphorylation
Kinase Assay	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.
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.
References	[1] https://pubmed.ncbi.nlm.nih.gov/9154836/ [2] https://pubmed.ncbi.nlm.nih.gov/24333448/ [3] https://pubmed.ncbi.nlm.nih.gov/22684030/ [4] https://pubmed.ncbi.nlm.nih.gov/23525555/

Applications

Methods	Biomarkers	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