Analysis Note

ControlA-549 cell lysate

Application

Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in human lymphoblastoid cell line (NL553) treated with NCS. (Khosravi, R., et al. (1999). PNAS. 96(26):14973–14977.)

Anti-MDM2 Antibody, clone 2A10 is a Mouse Monoclonal Antibody for detection of MDM2 also known as E3 ubiquitin-protein ligase Mdm2 or Hdm2 & has been validated in WB & IP.

General description

MDM2 is an E3 ubiquitin ligase that is most widely studied for its role in regulating the p53 tumor suppressor. MDM2 antagonizes the transcriptional activity of p53 in two main ways. The N-terminal of MDM2 interacts and disables the transactivation domain of p53. In addition, the C-terminal E3 ligase region of MDM2 can attach monoubiquitin to p53 resulting in the transport of p53 from the nucleus to the cytoplasm, thereby preventing the interaction of p53 with target DNA sequences. In addition, MDM2 may also attach polyubiquitin chains to p53, resulting in the degradation of p53 by the proteosome. MDM2 may produce these effects in cooperation with the Mdmx protein. Overall, MDM2 inhibits the ability of p53 to induce cell cycle arrest or apoptosis, and may contribute to oncogenesis.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Format: Purified

Purified mouse monoclonal IgG2aκ cultured supernatant in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Quality

Evaluated by Western Blot in A-549 cell lysate.

Western Blot Analysis: 0.5 µg/mL of this antibody detected MDM2 on 10 µg of A-549 cell lysate.

Target description

~90 kDa observed. The calculated molecular weight is 55 kDa, however MDM2 has been shown as a ~90 kDa band in western blots (Erhardt, P., et al. (1997). The Journal of Biological Chemistry. 272:15049-15052.). Uniprot describes many isoforms between 12-56 kDa, which can be highly phosphorylated and ubiquitinated.