UNC1215

Catalog No.S7088 Batch:S708801

Technical Data

Formula

C₃₂H₄₃N₅O₂

Molecular Weight

529.72

CAS No.

1415800-43-9

Solubility (25°C)*

In vitro

Ethanol

55 mg/mL (103.82 mM)

DMSO (warmed with 50ºC water bath)

20 mg/mL (37.75 mM)

Water

Insoluble

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

Clear solution

10% DMSO 1 40% PEG300 2 5%Tween80 3 45%ddH2O

2.0mg/ml

Taking the 1 mL working solution as an example, add 100 μL of 20 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 450 μL of ddH2O to adjust the volume to 1 mL. 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

UNC1215 is a potent and selective MBT (malignant brain tumor) antagonist, which binds L3MBTL3 with IC50 of 40 nM and K_d of 120 nM, 50-fold selective versus other members of the human MBT family.

Targets

L3MBTL3 ^[1]	L3MBTL3 ^[1]	L3MBTL3- D274A ^[1]
40 nM	120 nM(Kd)	3.5 μM

In vitro

UNC1215 binds L3MBTL3, competitively displacing mono- or dimethyllysine-containing peptides. This probe is greater than 50-fold selective versus other members of the human MBT family. UNC1215 has about a 75-fold selectivity for L3MBTL3 over L3MBTL1. UNC1215 shows no activity at concentrations up to 30 μM against the tandem Tudor domain of UHRF1, the chromodomain of CBX7 or the PHD domain of JARID1A. X-ray crystallography identifies a unique 2:2 polyvalent mode of interaction between UNC1215 and L3MBTL3. In cells, UNC1215 is nontoxic and directly binds L3MBTL3 via the Kme-binding pocket of the MBT domains. UNC1215 increases the cellular mobility of GFP-L3MBTL3 fusion proteins, and point mutants that disrupt the Kme-binding function of GFP-L3MBTL3 phenocopy the effects of UNC1215 on localization. UNC1215 is used to reveal a new Kme-dependent interaction of L3MBTL3 with BCLAF1, a protein implicated in DNA damage repair and apoptosis. ^[1]

Features

The first chemical probe for a Kme-binding protein.

Protocol (from reference)

Kinase Assay:^{^[1]}	AlphaScreen assay Compound plates (1 μL at 10 or 30 mM highest concentration) are diluted in 1×assay buffer (20 mM Tris pH 8.0, 25 mM NaCl, 2 mM DTT and 0.05% Tween-20) over 2 steps using a Multimek robotic pipettor and 1 μL is spotted into the wells of 384-well assay Proxiplates. To these plates 9 μL of protein- peptide mix in 1× assay buffer is added by Multidrop and incubated for 30 min at room temperature. At this point 2 μL of streptavidin-conjugate donor and nickel-chelate acceptor beads (45 μg/mL in 1× assay buffer) are added, the plates are allowed to incubate for an additional 30 min in the dark at room temperature. After incubation the plates are read on EnVision mulilabel reader equipped with HTS alpha screen laser. The screens reported are performed up to 10 or 30 μM, and therefore it should be noted that those compounds referred to as inactive are indeed inactive only within the concentration range tested. PHF23 and JARID1A are GST tagged and consequently for these assays GST-acceptor beads are used. It should be noted that any positive binding curves for L3MBTL4 that are generated yielded curves with very shallow slopes, suggesting a nonspecific interaction. The data for the IC50 values is calculated from replicate runs in that the datapoints for each compound concentration are averaged and plotted using 4-parameters curve fitting.
Cell Assay:^{^[1]}	Cell lines HEK293T/17 cells Concentrations ~100 μM Incubation Time 24 h Method CellTiter-Glo luminescent cell viability assay

Kinase Assay:^{^[1]}

AlphaScreen assay
Compound plates (1 μL at 10 or 30 mM highest concentration) are diluted in 1×assay buffer (20 mM Tris pH 8.0, 25 mM NaCl, 2 mM DTT and 0.05% Tween-20) over 2 steps using a Multimek robotic pipettor and 1 μL is spotted into the wells of 384-well assay Proxiplates. To these plates 9 μL of protein- peptide mix in 1× assay buffer is added by Multidrop and incubated for 30 min at room temperature. At this point 2 μL of streptavidin-conjugate donor and nickel-chelate acceptor beads (45 μg/mL in 1× assay buffer) are added, the plates are allowed to incubate for an additional 30 min in the dark at room temperature. After incubation the plates are read on EnVision mulilabel reader equipped with HTS alpha screen laser. The screens reported are performed up to 10 or 30 μM, and therefore it should be noted that those compounds referred to as inactive are indeed inactive only within the concentration range tested. PHF23 and JARID1A are GST tagged and consequently for these assays GST-acceptor beads are used. It should be noted that any positive binding curves for L3MBTL4 that are generated yielded curves with very shallow slopes, suggesting a nonspecific interaction. The data for the IC50 values is calculated from replicate runs in that the datapoints for each compound concentration are averaged and plotted using 4-parameters curve fitting.

Cell Assay:^{^[1]}

Cell lines
HEK293T/17 cells
Concentrations
~100 μM
Incubation Time
24 h
Method
CellTiter-Glo luminescent cell viability assay

References

[1] James LI, et al. Nat Chem Biol, 2013, 9(3), 184-191.

Selleck's UNC1215 has been cited by 8 publications

Recapitulating early human development with 8C-like cells [ Cell Rep, 2022, 39(12):110994]	PubMed: 35732112
Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer [ Cancers (Basel), 2022, 14(19)4883]	PubMed: 36230806
Peroxisome-driven ether-linked phospholipids biosynthesis is essential for ferroptosis [ Cell Death Differ, 2021, 28(8):2536-2551]	PubMed: 33731874
Peroxisome-driven ether-linked phospholipids biosynthesis is essential for ferroptosis [ Cell Death Differ, 2021, 10.1038/s41418-021-00769-0]	PubMed: 33731874
Prolonged unfolded protein reaction is involved in the induction of chronic myeloid leukemia cell death upon oprozomib treatment [ Cancer Sci, 2020, 112(1):133-143]	PubMed: 33067904
Epigenetic Reprogramming with Antisense Oligonucleotides Enhances the Effectiveness of Androgen Receptor Inhibition in Castration-Resistant Prostate Cancer [ Cancer Res, 2018, 78(20):5731-5740]	PubMed: 30135193
[ Cell Death Dis, 2018, ]	PubMed: 29988031
PHF20L1 antagonizes SOX2 proteolysis triggered by the MLL1/WDR5 complexes [Wang Q1, et al. Lab Invest, 2018, 98(12):1627-1641]	PubMed: 30089852

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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.

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