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Assay Method Information

Assay Name:  Fluorescence Polarization Cell-Free Homogeneous Dose Retest to Confirm Inhibitors of the LANA Histone H2A/H2B Interaction
Description:  Primary Collaborators: Kenneth Kaye,Brigham & Womens,Boston MA,kkaye@rics.bwh.harvard.edu,617-525-4256 Chantal Beauchemin,Brigham & Womens,Boston MA,cbeauchemin@rics.bwh.harvard.edu,617-525-4256 Keywords: KSHV, LANA, fluorescence polarization, nucleosomes, viral persistence Assay Overview: Screening for inhibitors that reduce or prevent the binding of LANA to the acidic region of the H2A/H2B histone dimer interface. Synthetic LANA peptide is labeled with an amino terminal FITC fluorophore linked via a beta alanine residue. The peptide contains the first 23 amino acids of the LANA protein that is essential for binding to the H2A/H2B interface. Nucleosomes were purifed from chicken Erythrocytes as a source of intact H2A/H2B dimers. Fluorescence is measured in the S and P planes with a Perkin Elmer Viewlux after 1 hour incubation. Expected Outcome: Compounds will be identified that inhibit the interaction of LANA 1-23 peptide with purified nucleosomes (containing H2A/H2B histone
Affinity data for this assay
 

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Last update November 1, 2007
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