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ML-323

Catalog No. T1757 CAS 1572414-83-5

Synonyms: ML323

ML323 is a reversible and effective USP1-UAF1 inhibitor in a Ub-Rho assay (IC50: 76 nM) and in orthogonal gel-based assays using K63-linked diubiquitin (di-Ub) (IC50: 174 nM 820 nM)and monoubiquitinated PCNA (Ub-PCNA) (IC50: 820 nM) as substrates, respectively.

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ML-323, CAS 1572414-83-5

Pack Size	Availability	Price/USD
5 mg	In stock	$ 52.00
10 mg	In stock	$ 77.00
25 mg	In stock	$ 102.00
50 mg	In stock	$ 158.00
100 mg	In stock	$ 239.00
1 mL * 10 mM (in DMSO)	In stock	$ 58.00

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Purity: 99.96%

Purity: 99.87%

Biological Description

Chemical Properties

Storage & Solubility Information

Description	ML323 is a reversible and effective USP1-UAF1 inhibitor in a Ub-Rho assay (IC50: 76 nM) and in orthogonal gel-based assays using K63-linked diubiquitin (di-Ub) (IC50: 174 nM 820 nM)and monoubiquitinated PCNA (Ub-PCNA) (IC50: 820 nM) as substrates, respectively.
Targets&IC50	USP1/UAF1:76 nM
In vivo	In H596 cells and U2OS osteosarcoma cells, ML323 targets two primary DNA damage response pathways (TLS and FA), enhancing the cytotoxicity of cisplatin. By inhibiting the USP1–UAF1 activity in H596 cells, ML323 suppresses the deubiquitination of PCNA and ANCD2.
Kinase Assay	High-throughput screening: For HTS, USP1-UAF1 activity is monitored using ubiquitin-rhodamine 110 as a substrate, where hydrolysis of the amide bond between the C-terminal glycine of ubiquitin and rhodamine results in an increase in fluorescence. The assay is miniaturized to a 4 μL volume in a 1,536-well format and is used to screen approximately 402,701 compounds in quantitative HTS mode, with each compound tested over a range of four to five concentrations. The assay shows robust performance with an average Z'factor of 0.8 throughout the screen.
Cell Research	For the colony-forming assay, cells are seeded at a density of 300–500 cells per well in six-well plates and grown overnight. Cells are then treated with ML323 alone, cisplatin alone or a combination of cisplatin and ML323 (1:1 or 1:4) at the indicated concentrations. Cells treated with an equal volume of DMSO and saline are used as control. After 48 h of treatment, fresh growth medium is added, and cells are incubated for an additional 5-10 d to allow for colony formation. For UV combination treatment, the cells are treated with ML323 at the indicated concentrations or an equal volume of DMSO. After 48 h, the medium is removed, and cells are irradiated at 254 nm at the indicated dosage. Fresh growth medium is added, and the cells are incubated for an additional 5-10 d to allow for colony formation. The cells without UV irradiation but treated with ML323 or an equal volume of DMSO are used as controls and designated as 100%. After the formation of the colonies, cells are fixed with methanol and stained with 0.5% crystal violet. Colonies consisting of >50 cells are scored. The number of colonies is determined from triplicate plates. The dose-response curves are generated using GraphPad Prism and analyzed by using CalcuSyn to calculate the combination index, which is determined for the fraction of cells affected after the addition of fixed ratios of cisplatin and the USP1-UAF1 inhibitor. (Only for Reference)

Synonyms	ML323
Molecular Weight	384.48
Formula	C23H24N6
CAS No.	1572414-83-5

Storage

Powder: -20°C for 3 years | In solvent: -80°C for 1 year

Solubility Information

Ethanol: 38.5 mg/mL (100 mM)

DMSO: 38.5 mg/mL (100 mM)

References and Literature

1. Liang Q, et al. Nat Chem Biol. 2014 , 10(4), 298-304.

Citations

1. Wu W, Xu H, Liao C, et al. Blockade of USP14 potentiates type I interferon signaling and radiation-induced antitumor immunity via preventing IRF3 deubiquitination. Cellular Oncology. 2022: 1-15 2. Yue X, Liu T, Wang X, et al.Pharmacological inhibition of BAP1 recruits HERC2 to competitively dissociate BRCA1–BARD1, suppresses DNA repair and sensitizes CRC to radiotherapy.Acta Pharmaceutica Sinica B.2023

Related compound libraries

This product is contained In the following compound libraries：

Inhibitor Library Anti-Cancer Compound Library Bioactive Compounds Library Max HIF-1 Signaling Pathway Compound Library NO PAINS Compound Library Anti-Aging Compound Library Cell Cycle Compound Library Bioactive Compound Library DNA Damage & Repair Compound Library Ubiquitination Compound Library

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ML364 XL177A Vialinin A USP7-IN-6 USP30 inhibitor 11 USP7-IN-8 IMP-1710 USP25/28 inhibitor AZ1

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Keywords

ML-323 1572414-83-5 Cell Cycle/Checkpoint DNA Damage/DNA Repair Ubiquitination DUB Deubiquitinase ML323 Inhibitor inhibit DUBs ML 323 inhibitor

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