| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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Purity: ≥98%
Mirin is a potent inhibitor of the MRN (Mre11 / Rad50 / Nbs1) complex and Ataxia Telangiectasia-Mutated (ATM) protein kinase. The Mre11/Rad50/Nbs1 (MRN) complex recruits the Ataxia Telangiectasia-Mutated (ATM) protein kinase to sites of double-strand DNA breaks and aids in ATM monomerization and activation. At least two different conformational states of MRN exist, and they are both reliant on ATP binding and Rad50 protein hydrolysis. Although some mutations in the Mre11 catalytic domain, as well as the mirin compound, block ATM activation independent of nuclease function, Mre11 nuclease activity is not necessary. Egg extracts and mammalian cells that have been treated with the Mre11 inhibitor mirin have less RCC1 binding to mitotic chromosomes. By modifying RCC1 chromosome association, the MRN-CtIP pathway thereby aids in Ran-dependent mitotic spindle assembly.
| ln Vitro |
Mirin inhibits the phosphorylation of Nbs1 and Chk2 downstream targets by ATM, the DSB-induced activation of ATM, and the MRN-dependent autophosphorylation of ATM at Ser1981 in response to DSBs. Additionally, mirin prevents homology-dependent DNA repair in HEK293 cells and the G2 checkpoint in TOSA4 cells.[1]
Mirin makes HPV episomes more sensitive to PA25 in cells that have integrated HPV16 (SiHa), which lowers the PA25 IC50 by about five times.[2] In addition, pretreatment with mirin reduces the viability of the cells and prevents the expression of the nuclear antigen in proliferating cells in human embryonic kidney 293 cells treated with cisplatin.[3] |
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| Enzyme Assay |
In 25 mM MOPS (pH 7.0), 60 mM KCl, 0.2% Tween 20, 2 mM DTT, 1 mM or 5 MnCl2 (or 5 mM MgCl2, or 5 mM CaCl2), 0.1 pmol of DNA substrate, and 0.3 pmol of Mre11 (or an equivalent amount of Mre11 complexed with Rad50) in a volume of 10 μl are the contents of reactions with oligonucleotide nonhairpin substrates. The reactions are then incubated at 37°C for 30 minutes. After that, proteinase K, SDS, and EDTA are added at final concentrations of 0.1 mg/ml, 5 mM, and 0.2%, respectively, and the mixture is incubated for an additional 15 minutes. A sequencing gel containing 10% acrylamide and 7 M urea is loaded with 4 μl of each reaction combined with 4 μl of formamide loading buffer. Each gel is examined using a phosphorimaging apparatus following the run. The only differences between reactions with and without hairpin substrates are that 3 pmol of Mre11 is added to the reactions as directed and that the reactions are left to overnight at room temperature. 25 mM MOPS (pH 7.0), 60 mM KCl, and 0.2% Tween are included in nonhomologous end-joining reactions. 0.2 mM DTT, 0.5 mM ATP, 4 ng of plasmid DNA, 10% polyethylene glycol, 0.01 pmol of human DNA ligase I, 0.06 pmol of Mre11, and 0.1 units of E. coli exonuclease III (GIBCO-BRL) in a volume of 10 μl are all included in the mixture. Following a 25-minute incubation period at 37°C, a final concentration of 0.5% of Tween 20 is added, and a 2.5-microliter aliquot is amplified via PCR using primers DAR5 and DAR147. The TA cloning kit is used to clone PCR products, and an automated ABI Capillary Genetic Analyzer is used to sequence the results.
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| Cell Assay |
In humidified air with 5% CO2 at 37 °C, human embryonic kidney (HEK) 293 cells are cultured in RPMI-1640 supplemented with 5% heat-inactivated fetal bovine serum, penicillin (100 U/mL), and streptomycin (100 mg/mL). Every 48 hours, new medium is added to the cells. In 96-well plates, the cells are seeded with regular growth medium. Before receiving cisplatin (20 μM) treatment and being incubated for eight and twenty-four hours, cells are pretreated with mirin (100 μM) for one hour. Following the manufacturer's instructions, the MTT assay is carried out using the EZ-Cytox cell viability assay kit, and MTT reduction is measured using a micro-plate reader at a wavelength of 450 nm.
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| References |
| Molecular Formula |
C10H8N2O2
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| Molecular Weight |
220.25
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| Exact Mass |
220.03
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| Elemental Analysis |
C, 54.53; H, 3.66; N, 12.72; O, 14.53; S, 14.56
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| CAS # |
1198097-97-0
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| Related CAS # |
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| Appearance |
Solid powder
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| SMILES |
C1=CC(=CC=C1/C=C\2/C(=O)NC(=N)S2)O
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| InChi Key |
YBHQCJILTOVLHD-YVMONPNESA-N
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| InChi Code |
InChI=1S/C10H8N2O2S/c11-10-12-9(14)8(15-10)5-6-1-3-7(13)4-2-6/h1-5,13H,(H2,11,12,14)/b8-5-
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| Chemical Name |
(5Z)-5-[(4-hydroxyphenyl)methylidene]-2-imino-1,3-thiazolidin-4-one
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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) |
DMSO: ~44 mg/mL (~199.8 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (4.54 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1 mg/mL (4.54 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 1 mg/mL (4.54 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.5403 mL | 22.7015 mL | 45.4030 mL | |
| 5 mM | 0.9081 mL | 4.5403 mL | 9.0806 mL | |
| 10 mM | 0.4540 mL | 2.2701 mL | 4.5403 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.
Inhibition of MRN results in prolonged metaphase in mammalian cells.Mol Cell.2013 Mar 28;49(6):1097-107. |
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MRN inhibition disrupts the RanGTP gradient during metaphase.Mol Cell.2013 Mar 28;49(6):1097-107. |
MRN inhibition results in a reduction of RCC1 binding to chromatin.Mol Cell.2013 Mar 28;49(6):1097-107. |
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