RI-1

Catalog No.S8077 Synonyms: RAD51 inhibitor 1

For research use only.

RI-1 (RAD51 inhibitor 1) is a RAD51 inhibitor with IC50 ranging from 5 to 30 μM.

RI-1 Chemical Structure

CAS No. 415713-60-9

Selleck's RI-1 has been cited by 15 publications

Purity & Quality Control

Choose Selective RAD51 Inhibitors

Biological Activity

Description RI-1 (RAD51 inhibitor 1) is a RAD51 inhibitor with IC50 ranging from 5 to 30 μM.
Features A selective recombinant RAD51 protein inhibitor discovered in 2012. Valuable tool for mechanistic studies of DNA repair and potential for use in many cancers.
Targets
RAD51 [1]
<30 μM
In vitro

RI-1 sensitizes cells to DNA damage by directly and specifically disrupting HsRAD51 and inhibiting the ability of RAD51 to form filaments on ssDNA. In addition, RI-1 alone generates single-agent toxicity in all three cancer cell lines (HeLa, MCF-7 and U2OS) with LD50 values in the 20–40 µM range. [1] RI-1 decreases the rejoining of γ-H2AX foci in G2 phase cells and results in a higher level of unrepaired DSBs 6 hours after irradiation. [2]

Protocol (from reference)

Kinase Assay:[1]
  • DNA binding assays:

    All reactions are performed in black non-binding polystyrene 384-well plates with reaction volumes of 30–100 μL. Purified DNA strand exchange proteins and chemical compounds are pre-incubated at room temperature for 5 minutes; they are then further incubated at 37°C for 30 min with 100 nM of ssDNA substrate, consisting of a 45-mer poly-dT tagged with Alexa 488 at the 5’ terminus (synthesized and purified by Integrated DNA Technologies). Reactions are performed in 20 mM HEPES pH 7.5, 10 mM MgCl2, 0.25 μM BSA, 2% glycerol, 30 mM NaCl, 4% DMSO and 2 mM ATP. Some conditions included DTT or TCEP (tris(2-carboxyethyl)phosphine) as indicated. DNA binding is measured as a function of fluorescence polarization (FP) with a Safire2 plate reader, using the following settings: excitation 470±5nm, emission 530±5nm, 10 reads/well, Z height and G factor auto-calibrated from control wells. Displayed error bars represent standard deviation. For experiments involving a titration of protein concentrations, data are fit to an equation that accounts for the cooperative nature by which recombinase proteins bind DNA. For experiments involving a titration of RI-1, protein concentrations are selected to give an ∼80% saturation of the FP signal in the absence of RI-1.

Cell Research:[1]
  • Cell lines: HeLa, MCF-7 and U2OS cells
  • Concentrations: ~35 μM
  • Incubation Time: 24 hours
  • Method: Cytotoxicity is determined by loss of colony-forming ability. Experiments are performed in triplicate. Crystal violet stained colonies are imaged with a CCD camera and counted using NIH Image software. Error bars denote standard error.

Solubility (25°C)

In vitro

Chemical Information

Molecular Weight 361.61
Formula

C14H11Cl3N2O3

CAS No. 415713-60-9
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles C1COCCN1C2=C(C(=O)N(C2=O)C3=CC(=C(C=C3)Cl)Cl)Cl

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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.

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Molarity Calculator

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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