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67649673450
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note
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ELISA assay: The procedure in Ref. 16 was followed. Briefly, His6-p53 was immobilized on 96-well ELISA plates. GST-MDM2 was then added in the absence or presence of potential inhibitors. GST-MDM2 bound to p53 was detected using MDM2 monoclonal antibody 5B10, followed by incubation with protein A-HRP conjugate and a chromogenic substrate.
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23
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67649664856
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note
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The AlphaScreen™ assay, which is available from Perkin Elmer, relies upon donor and acceptor beads that are coated with hydrogel to provide functional groups for bioconjugation. When two proteins interact (one of which is conjugated to the donor bead and the other to the acceptor bead) the beads are brought into close proximity. Excitation of a photosensitizer in the donor bead results in the production of singlet oxygen that diffuses to and reacts with a chemiluminescent molecule in the acceptor bead. This results in the activation of fluorophores in the acceptor bead which results emitted light that can be detected.
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24
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67649640293
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note
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50 values reported are from runs repeated four times.
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67649662888
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note
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+) 353.0831, found 353.0829.
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37649000332
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Betzi S., Restouin A., Opi S., Arold S.T., Parrot I., Guerlesquin F., Morelli X., and Collette Y. Proc. Natl. Acad. Sci. U.S.A. 104 (2007) 19256
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Morelli, X.7
Collette, Y.8
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