The Scripps Research Institute

Ventana Medical Systems, Inc.

Xiao-Bo Chen

Christopher Bieniarz

Michael Patrick Farrell

Paul R Russell

Michael N Boddy

Clare H McGowan

Veronique Blais

Pierre-Henri L Gaillard

Disclosed embodiments concern quantifying a biomolecule conjugated to a nanoparticle. Quantifying typically comprises determining the number of biomolecules per nanoparticle. Any suitable biomolecule can be used, including but not limited to, amino acids, peptides, proteins, haptens, nucleic acids, oligonucleotides, DNA, RNA, and combinations thereof. A single type of biomolecule may be conjugated to the nanoparticle, more than one biomolecule of a particular class may be conjugated to the nanoparticle, or two or more classes of biomolecules may be conjugated to the nanoparticle. Certain disclosed embodiments comprise enzymatically or chemically digesting a biomolecule conjugated to the nanoparticle, or displacing a biomolecule using ligand-exchange chemistry. Where biomolecule concentrations are determined, any technique suitable for determining biomolecule concentration can be used, such as spectrophotometric techniques, including measuring tryptophan fluorescence and using a standard fluorescence intensity versus biomolecule concentration curve.

Method for quantifying biomolecules conjugated to a nanoparticle

Isolated mammalian Mus81-Eme endonuclease complexes comprise an Mus81 protein portion and an Eme protein portion. A method of identifying a chemical compound that modulates mammalian cellular response to DNA damage comprises contacting a chemical compound to be tested with a biochemical mixture containing an isolated mammalian Mus81-Eme1 endonuclease complex, a source of magnesium ion, and a suitable DNA substrate; measuring the activity level of mammalian Mus81-Eme endonuclease complex in the mixture; comparing the measured activity level to the activity level of a substantially similar mixture of isolated Mus81-Eme1 endonuclease, magnesium ion, and DNA substrate in the absence of the chemical compound to be tested; and selecting a chemical compound that increases or decreases the endonuclease activity. Isolated mammalian Eme1 and Eme2 proteins derived from humans and murine species and isolated nucleic acids encoding the proteins are also described.

Mammalian endonucleases and methods of use

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Tucson, AZ, United States of America

Xiao-Bo Chen