Phospho-MDM2-S166 Rabbit mAb for IHC-P and ELISA - Q00987

Core Product Specifications and Parameters

Biological Background: MDM2 Function and Localization

• MDM2 is an E3 ubiquitin-protein ligase that mediates ubiquitination of p53/TP53, leading to its proteasomal degradation and inhibition of p53- and p73/TP73-mediated cell cycle arrest and apoptosis.
  Related references:
  PMID:29681526
• It also ubiquitinates and promotes the degradation of other tumor suppressors and signaling proteins, including RB1, DAXX, IGF1R, and SNAI1.
  Related references:
  PMID:12821780
  PMID:15053880
  PMID:15195100
• MDM2 negatively regulates mitochondrial respiration by binding NDUFS1, causing its cytosolic retention, reduced complex I activity, oxidative stress, and apoptosis.
  Related references:
  PMID:30879903
• Predominantly localized to the nucleus (nucleoplasm and nucleolus) with additional cytoplasmic distribution.
• Ubiquitously expressed in normal tissues; cancer-specific isoforms (Mdm2-A through Mdm2-G) are detected in a range of malignancies.
• Phosphorylation at serine 166 and other residues regulates MDM2 activity and protein-protein interactions.
• Contains a RING-type zinc finger domain and belongs to the ubiquitin-conjugating enzyme family, requiring zinc for its E3 ligase function.

Experimental Guidance and Technical Tips

• The antibody was raised against a phospho-synthetic peptide surrounding S166; it specifically recognizes the phosphorylated form of MDM2. Validate the phosphorylated status with phosphatase treatment controls.
• For IHC-P, consider antigen retrieval (e.g., citrate buffer) and validate staining intensity in tissue samples known to harbor elevated phospho-MDM2 levels (e.g., certain tumors).
• For ELISA, optimize coating conditions and antibody dilution by checkerboard titration, and include both phosphorylated and non-phosphorylated peptide controls to confirm specificity.
• Cross-reactivity with mouse and rat has been reported; verify signal in the relevant species’ samples using appropriate positive and negative controls.

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