Nutlin-3b

Catalog No.S8065 Batch:S806501

Technical Data

Formula	C₃₀H₃₀Cl₂N₄O₄
Molecular Weight	581.49	CAS No.	675576-97-3
Solubility (25°C)*	In vitro	DMSO	100 mg/mL (171.97 mM)
		Ethanol	100 mg/mL (171.97 mM)
		Water	Insoluble
* <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

Nutlin-3b ((+)-Nutlin-3) is a p53/MDM2 antagonist or inhibitor with IC50 value of 13.6 μM, 150-fold less potent (+)-enantiomer of Nutlin-3 as in comparison with opposite (-)-enantiomer Nutlin-3a.

Targets

MDM2 ^[1]

13.6 μM

In vitro

Nutlin-3b is useful as a negative control s for non-MDM2-related cellular activities. Nutlin-3a induces the expression of MDM2 and p21 (but not p53) only in cells with wild-type p53. Nutlin-3b has no effect regardless of the p53 status of the cells. Only the active enantiomer Nutlin-3a shows a potent antiproliferative activity and clear separation of potency between the cells harboring wild-type p53 and those harboring mutant p53. The potency of Nutlin-3b is much lower in the wild-type p53 cells and nearly identical to the potency of Nutlin-3a against the mutant p53 cells. After 48 hours of exposure to the Nutlin-3a, 45% of the cell population became TUNEL positive, but cells treated with the Nutlin-3b are indistinguishable from the untreated controls. ^[1]

Protocol (from reference)

Kinase Assay:^{^[1]}	Biacore study Competition assay is performed on a Biacore S51. A Series S Sensor chip CM5 is utilized for the immobilization of a PentaHis antibody for capture of the His-tagged p53. The level of capture is ~200 response units (1 response unit corresponds to 1 pg of protein per mm ²). The concentration of MDM2 protein is kept constant at 300 nM. Nutlin-3 is dissolved in DMSO at 10 mM and further diluted to make a concentration series of Nutlin-3 in each MDM2 test sample. The assay is run at 25°C in running buffer (10 mM Hepes, 0.15 M NaCl, 2% DMSO). MDM2-p53 binding in the presence of Nutlin-3 is calculated as a percentage of binding in the absence of Nutlin-3 and IC50 is calculated
Cell Assay:^{^[1]}	Cell lines HCT116, RKO, SJSA-1, SW480, and MDA-MB-435 Concentrations ~30 μM Incubation Time 48 h Method MTT assay

Kinase Assay:^{^[1]}

Biacore study
Competition assay is performed on a Biacore S51. A Series S Sensor chip CM5 is utilized for the immobilization of a PentaHis antibody for capture of the His-tagged p53. The level of capture is ~200 response units (1 response unit corresponds to 1 pg of protein per mm ²). The concentration of MDM2 protein is kept constant at 300 nM. Nutlin-3 is dissolved in DMSO at 10 mM and further diluted to make a concentration series of Nutlin-3 in each MDM2 test sample. The assay is run at 25°C in running buffer (10 mM Hepes, 0.15 M NaCl, 2% DMSO). MDM2-p53 binding in the presence of Nutlin-3 is calculated as a percentage of binding in the absence of Nutlin-3 and IC50 is calculated

Cell Assay:^{^[1]}

Cell lines
HCT116, RKO, SJSA-1, SW480, and MDA-MB-435
Concentrations
~30 μM
Incubation Time
48 h
Method
MTT assay

References

[1] Vassilev LT, et al. Science, 2004, 303(5659), 844-848.

Customer Product Validation

: Data from [Data independently produced by , , Electrophoresis, 2015, 36(24):3101-4]

Selleck's Nutlin-3b has been cited by 2 publications

Nucleotide biosynthesis links glutathione metabolism to ferroptosis sensitivity [ Life Sci Alliance, 2022, 5(4)e202101157]	PubMed: 35074928
Nondenaturing polyacrylamide gel electrophoresis to study the dissociation of the p53·MDM2/X complex by potentially anticancer compounds. [Sgammato R, et al. Electrophoresis, 2015, 36(24):3101-4]	PubMed: 26383830

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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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