RG108

Catalog No.S2821 Batch:S282104

Technical Data

Formula

C₁₉H₁₄N₂O₄

Molecular Weight

334.33

CAS No.

48208-26-0

Solubility (25°C)*

In vitro

DMSO

67 mg/mL (200.4 mM)

Ethanol

67 mg/mL (200.4 mM)

Water

Insoluble

In vivo (Add solvents to the product individually and in order)

Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	3.35mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 67 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	0.415mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 8.3 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.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

RG108 (N-Phthalyl-L-tryptophan) is an inhibitor of DNA methyltransferase with IC50 of 115 nM in a cell-free assay, does not cause trapping of covalent enzymes.

Targets

DNA methyltransferase ^[1]
(Cell-free assay)

115 nM

In vitro

RG108 effectively blocks DNA methyltransferases in vitro and does not cause covalent enzyme trapping in human cell lines. Incubation of cells with low micromolar concentrations of RG108 results in significant demethylation of genomic DNA without any detectable toxicity. Intriguingly, RG108 causes demethylation and reactivation of tumor suppressor genes, but it does not affect the methylation of centromeric satellite sequences. ^[1] In another study, the synthesis and in vitro analysis of a biotinylated RG108 conjugate is investigated to evaluate the interactions with DNA methyltransferase enzymes. ^[2] In a recent study, it is shown RG108 can significantly reduce the DNA methyltransferases activity in SM derived iPS cells as compared to the native SMs. ^[3]

Protocol (from reference)

Kinase Assay:^{^[1]}	In vitro methylation assay The substrate DNA for the in vitro methylation assay is a 798 bp fragment (−423/+375 relative to the initiation codon) from the promoter region of the human p16Ink4a gene. The methylation reaction contains 350 to 400 ng substrate DNA and 4 units of M.SssI methylase (0.5 μM) in a final volume of 50 μL. Inhibitors are added to final concentrations of 10, 100, 200, and 500 μM, respectively. Reactions are done at 37 °C for 2 hours. After completion, the reaction is inactivated at 65 °C for 15 minutes and the DNA is purified using PCR Purification kit. Three hundred nanograms of purified DNA is digested for 3 hours at 60 °C with 30 units of BstUI and analyzed on 2% Tris-borate EDTA agarose gels.
Cell Assay:^{^[1]}	Cell lines HCT116 cells Concentrations 1-100 μM Incubation Time 5 days Method For the determination of cellular growth and viability, cells are stained with trypan blue and counted using a standard counting grid.

Kinase Assay:^{^[1]}

In vitro methylation assay
The substrate DNA for the in vitro methylation assay is a 798 bp fragment (−423/+375 relative to the initiation codon) from the promoter region of the human p16Ink4a gene. The methylation reaction contains 350 to 400 ng substrate DNA and 4 units of M.SssI methylase (0.5 μM) in a final volume of 50 μL. Inhibitors are added to final concentrations of 10, 100, 200, and 500 μM, respectively. Reactions are done at 37 °C for 2 hours. After completion, the reaction is inactivated at 65 °C for 15 minutes and the DNA is purified using PCR Purification kit. Three hundred nanograms of purified DNA is digested for 3 hours at 60 °C with 30 units of BstUI and analyzed on 2% Tris-borate EDTA agarose gels.

Cell Assay:^{^[1]}

Cell lines
HCT116 cells
Concentrations
1-100 μM
Incubation Time
5 days
Method
For the determination of cellular growth and viability, cells are stained with trypan blue and counted using a standard counting grid.

References

Customer Product Validation

: Data from [Data independently produced by Biosens Bioelectron, 2015, 66, 109-14]

: Data from [Data independently produced by , , Cell Physiol Biochem, 2018, 50(4):1376-1397]

: Data from [Data independently produced by , , Oncotarget, 2015, 6(7):5134-46.]

: Data from [Data independently produced by , , Oncol Rep, 2018, 39(3):993-1002]

Selleck's RG108 has been cited by 27 publications

Association of DNA methylation/demethylation with the functional outcome of stroke in a hyperinflammatory state [ Neural Regen Res, 2024, 19(10):2229-2239]	PubMed: 38488557
Association of DNA methylation/demethylation with the functional outcome of stroke in a hyperinflammatory state [ Neural Regeneration Research, 2024, 19(10):p 2229-2239]	PubMed: none
Deciphering the potential ability of RG108 in cisplatin-induced HEI-OC1 ototoxicity: a research based on RNA-seq and molecular biology experiment [ Hereditas, 2023, 160(1):18]	PubMed: 37088824
Direct chemical reprogramming of human cord blood erythroblasts to induced megakaryocytes that produce platelets [ Cell Stem Cell, 2022, 29(8):1229-1245.e7]	PubMed: 35931032
Inhibiting DNA methylation alleviates cisplatin-induced hearing loss by decreasing oxidative stress-induced mitochondria-dependent apoptosis via the LRP1-PI3K/AKT pathway [ Acta Pharm Sin B, 2022, 12(3):1305-1321]	PubMed: 35530135
Recapitulating early human development with 8C-like cells [ Cell Rep, 2022, 39(12):110994]	PubMed: 35732112
Drug screen in iPSC-Neurons identifies nucleoside analogs as inhibitors of (G4C2)n expression in C9orf72 ALS/FTD [ Cell Rep, 2022, 39(10):110913]	PubMed: 35675776
Comprehensive drug response profiling and pan-omic analysis identified therapeutic candidates and prognostic biomarkers for Asian cholangiocarcinoma [ iScience, 2022, 25(10):105182]	PubMed: 36248745
Direct Reprograming of Mouse Fibroblasts into Dermal Papilla Cells via Small Molecules [ Int J Mol Sci, 2022, 23(8)4213]	PubMed: 35457029
The DNA methylation inhibitor RG108 protects against noise-induced hearing loss [ Cell Biol Toxicol, 2021, 10.1007/s10565-021-09596-y]	PubMed: 33723744

RETURN POLICY
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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.