Nutlin-3a, IC-03346914
Product Introduction
Nutlin-3a is the active enantiomer of Nutlin-3, a potent, cell-permeable, and selective small-molecule inhibitor of the murine double minute 2 (MDM2) protein. Nutlin-3a binds specifically to the p53-binding pocket of MDM2 with high affinity (IC50 ¡Ö 90 nM), thereby preventing MDM2-mediated ubiquitination and proteasomal degradation of the tumor suppressor protein p53. By stabilizing and activating p53, Nutlin-3a induces p53-dependent transcriptional programs leading to cell cycle arrest, apoptosis, and senescence in cancer cells harboring wild-type p53 while sparing p53-mutant or p53-null cells. Nutlin-3a is extensively utilized in cancer biology and pharmacology research to dissect the p53-MDM2 regulatory axis, investigate p53-dependent cellular responses, and evaluate MDM2 as a therapeutic target. Its high selectivity for MDM2 over MDMX (MDM4) and other p53-interacting proteins makes it a valuable pharmacological tool for specifically interrogating the MDM2-p53 interaction.
Product Features
1. Potent and Selective MDM2 Inhibitor: Displaces p53 from MDM2 with an IC50 of approximately 90 nM, activating p53-dependent transcription and tumor suppression.
2. Active Enantiomer: Supplied as the biologically active Nutlin-3a enantiomer, providing significantly higher potency than the racemic mixture or inactive enantiomer.
3. p53-Dependent Activity: Selectively induces cell cycle arrest, apoptosis, and senescence in cancer cells expressing wild-type p53, with minimal effects on p53-mutant or p53-null cells.
4. Extensive Preclinical Validation: Widely cited in cancer research for studying p53 signaling, MDM2 biology, and chemosensitization in both in vitro and in vivo models.
5. High Purity: Supplied at 99% purity for reproducible pharmacological studies.
Specifications
Size: Available in 5 mg solid, 1 mL (10 mM in DMSO), 10 mg, 25 mg, 50 mg, and 100 mg
CAS Number: 675576-98-4
Purity: 99%
Molecular Weight: 581.49
Molecular Formula: C30H30Cl2N4O4
Solubility: DMSO: ¡Ý30 mg/mL (stock solutions); limited solubility in aqueous buffers
Storage and Stability
Storage Conditions: Store sealed in dry conditions at room temperature (15-25¡ãC). Protect from light and moisture. For the 10 mM DMSO solution, store at -20¡ãC protected from light.
Shelf Life: The product is stable for 24 months from the date of manufacture when stored as directed. For reconstituted solutions in DMSO, aliquot into single-use volumes and store at -20¡ãC or -80¡ãC. Avoid repeated freeze-thaw cycles.
Protocol (For Reference Only)
Important: Nutlin-3a is the active enantiomer; verify enantiomer identity before use. Protect from light and moisture during handling and storage. Prepare stock solutions in anhydrous DMSO.
1. Stock Solution Preparation (Solid): Dissolve Nutlin-3a solid in anhydrous DMSO to prepare a concentrated stock solution (e.g., 10-50 mM). Aliquot into single-use vials and store at -20¡ãC protected from light. The 10 mM ready-to-use DMSO solution may be used directly for dilution into culture medium.
2. In Vitro Working Solution Preparation: Dilute DMSO stock solution into pre-warmed complete culture medium to achieve working concentrations typically ranging from 1-10 µM. Limit final DMSO concentration in culture medium to ¡Ü0.1% to avoid solvent-related cytotoxicity.
3. p53 Activation Assay: For p53 stabilization and activation, treat wild-type p53-expressing cells with Nutlin-3a at 2-10 µM for 6-24 hours. Assess p53 protein accumulation by Western blot, and evaluate transcriptional activation of p53 target genes (e.g., p21, MDM2, PUMA, BAX) by qPCR or Western blot.
4. Cell Cycle and Apoptosis Analysis: Treat cells with Nutlin-3a at 2-10 µM for 24-72 hours and assess cell cycle arrest by propidium iodide staining with flow cytometry, apoptosis by annexin V/PI dual staining, and cell viability by MTT or clonogenic survival assays.
5. Senescence Induction: For p53-dependent senescence studies, treat cells with Nutlin-3a at 5-10 µM for 5-7 days and evaluate senescence-associated ¦Â-galactosidase (SA-¦Â-gal) activity, morphological changes, and senescence-associated secretory phenotype (SASP) markers.
Precautions
1. Verify p53 status of experimental cell lines before use; Nutlin-3a specifically activates wild-type p53 and is ineffective in p53-mutant or p53-null cells.
2. Protect from light and moisture during handling and storage; Nutlin-3a is hygroscopic and photodegradable.
3. Use appropriate vehicle controls in all experiments; DMSO concentration in culture medium should not exceed 0.1% to avoid solvent-related cytotoxicity.
4. Nutlin-3a exhibits differential potency across cell types depending on p53 expression levels, MDM2 amplification status, and downstream pathway integrity; dose-response optimization is recommended for specific experimental systems.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the difference between Nutlin-3 and Nutlin-3a?
A1: Nutlin-3 is a racemic mixture of two enantiomers, while Nutlin-3a is the purified active enantiomer. Nutlin-3a is approximately 150-fold more potent than its inactive counterpart Nutlin-3b. Always verify that the product specified is the active Nutlin-3a enantiomer for functional studies.
Q2: What is the recommended concentration for p53 activation?
A2: Nutlin-3a at 2-10 µM effectively stabilizes and activates p53 in most wild-type p53-expressing cell lines. For complete MDM2 inhibition and maximal p53 induction, 5-10 µM is commonly used. Titration experiments are recommended for specific cell types.
Q3: Does Nutlin-3a affect cells with mutant p53?
A3: Nutlin-3a is highly selective for wild-type p53 and does not stabilize or activate mutant p53. It may exhibit limited off-target effects at high concentrations (>20 µM) in p53-mutant cells, but these effects are generally p53-independent. Verify p53 status in experimental cell lines before interpreting results.
Q4: Can Nutlin-3a be used in vivo?
A4: Yes. Nutlin-3a has been administered in rodent xenograft models via oral gavage or intraperitoneal injection at doses ranging from 100-400 mg/kg. It demonstrates p53-dependent antitumor activity in wild-type p53 xenografts. Due to species differences in MDM2 binding, Nutlin-3a exhibits reduced potency in mouse cells compared to human cells.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of conditions using a small-scale pilot experiment is strongly recommended before processing valuable samples.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-03346914
Product Name: Nutlin-3a
Size: 5 mg / 1 mL (10 mM in DMSO) / 10 mg / 25 mg / 50 mg
Price:
5 mg: CNY ¥1650.00 / USD $165.00 / EUR €198.00 / JPY ¥29700.00
1 mL (10 mM in DMSO): CNY ¥1900.00 / USD $190.00 / EUR €228.00 / JPY ¥34200.00
10 mg: CNY ¥1950.00 / USD $195.00 / EUR €234.00 / JPY ¥35100.00
25 mg: CNY ¥3200.00 / USD $320.00 / EUR €384.00 / JPY ¥57600.00
50 mg: CNY ¥4800.00 / USD $480.00 / EUR €576.00 / JPY ¥86400.00
2023 Version
