Products from BPS Bioscience require a minimum order value above 400€Application: Study binding and screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.
Background: TP53 encodes transcription factor p53, a tumor suppressor protein that maintains genomic stability and has been termed the "guardian of the genome". It is activated in response to cellular stress, such as DNA damage, oxidative stress, or oncogene activation. p53 regulates the expression of genes involved in repairing damaged DNA, preventing mutations from accumulating. It induces cell cycle arrest (commonly at the G1/S checkpoint) to allow DNA repair before cell division, and it induces apoptosis if damage is irreparable. In addition, it can regulate senescence to prevent the proliferation of damaged cells. TP53 is one of the most frequently mutated genes in human cancers, found in over 50% of cases. Mutations often result in loss of normal p53 function, although gain-of-function mutations are also observed, leading to generally more aggressive cancer phenotypes. TP53 mutation correlates with increased tumor growth, with resistance to chemotherapy, and with increased risk of metastasis. p53 protein is regulated by its interaction with MDM2 (mouse double minute 2 homolog), which serves as a ubiquitin ligase (E3) to target p53 for degradation. MDM2 ubiquitinates p53, resulting in the rapid degradation of p53 through the Ub–proteasome pathway. MDM2-mediated destabilization and inactivation of p53 plays a critical role in several human cancers. The disruption of the MDM2-p53 interaction has been regarded as an attractive strategy for anticancer drug discovery, as it would restore the tumor suppressor function of p53.
Contraindications: The MDM2-p53 Homogenous Assay Kit is compatible with up to 1% final DMSO concentration.Avoid green and blue dyes that absorb light in the AlphaScreen signal emission range (λ=520-620 nm), such as Trypan Blue.Avoid the use of potent singlet oxygen quenchers such as sodium azide (NaN
3) or metal ions (Fe
2+, Fe
3+, Cu
2+, Zn
2+ and Ni
2+).The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. Media like MEM, which lack these components, do not affect AlphaScreen assays.
Description: The MDM2-p53 Homogenous Assay Kit is designed to identify and assess the efficacy of inhibitors of MDM2 (mouse double minute 2 homolog) interaction with p53 by measuring their binding. The MDM2-p53 Homogeneous Assay Kit comes in a convenient 384-well AlphaLISA® format, with enough purified recombinant MDM2, p53, and buffers for 400 enzyme reactions.
Figure 1: Illustration of the assay principle. MDM2 contains a GST-tag, recognized by the GSH (glutathione) donor bead, while p53 contains a FLAG-tag that binds to the AlphaLISA™ acceptor bead conjugated with an anti-FLAG antibody. Upon excitation of the donor bead, a singlet oxygen is generated by the donor bead, which excites the acceptor bead and emits light proportionally to the level of MDM2-p53 interaction. AlphaLISA™ immunoassays are a no-wash alternative to ELISAs.Need us to run inhibitor screens or profile your compounds against MDM2 binding to p53? Check out our
Ubiquitination Screening Services. Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Uniprot: Q00987, Q29537
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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