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Active Mdm2 p53 Binding Protein Homolog (MDM2)

HDMX; hdm2; Mouse Double Minute 2,Human Homolog Of p53-Binding Protein; Oncoprotein Mdm2

ACTIVITY TEST

Mouse double minute 2 homolog (MDM2) also known as E3 ubiquitin-protein ligase. MdM2 is a cellular oncoprotein that recognizes the N-terminal trans-activation domain (TAD) of the p53 tumor suppressor and as an inhibitor of p53 transcriptional activation. The human homologue of this protein is sometimes called Hdm2. The p53 tumor suppressor is the key target of MDM2. It has been identified as a p53 interacting protein that represses p53 transcriptional activity and also acts as an E3 ubiquitin ligase, targeting both itself and p53 for degradation by the proteasome. MDM2 is capable of auto-polyubiquitination, and in complex with p300, a cooperating E3 ubiquitin ligase, is capable of polyubiquitinating p53. Besides, S100 Calcium Binding Protein (S100) has been identified as an interactor of MDM2, thus a binding ELISA assay was conducted to detect the interaction of recombinant human MDM2 and recombinant human S100. Briefly, MDM2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to S100-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MDM2 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution , wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450 nm immediately. The binding activity of recombinant human MDM2 and recombinant human S100 was shown in Figure 1, the EC50 is 0.037 ug/ml.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Clin Cancer Res Inactivation of the p53–KLF4–CEBPA Axis in Acute Myeloid Leukemia PubMed: 26408402
Clin Cancer Res Inactivation of the p53–KLF6–CEBPA Axis in Acute Myeloid Leukemia PubMed: 26408402
Analyst Sensitive and simultaneous surface plasmon resonance detection of free and p53-bound MDM2 proteins from human sarcomas Pubmed:29637949
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