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US 9,631,226 B2Granted
Agents providing controls and standards for immunoprecipitation assays
AssigneeThe General Hospital Corporation
InventorsBradley E. Bernstein, Alon Goren
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Abstract
Control agents for immunoprecipitation assays, methods of using the control agents and kits comprising the control agents are provided.
Claims16
A method of validating a chromatin immunoprecipitation (ChIP) assay result, the method comprising:
- (a) obtaining an input sample of the genomic material of interest and of one or more polypeptide-oligonucleotide conjugate of the formula:
- A-L-N,
- wherein A is a polypeptide comprising 5 amino acids, L is a linker and N is an oligonucleotide comprising 10 nucleotides, wherein the sequence of the 10 nucleotides of N uniquely identifies an amino acid sequence and/or amino acid modification of A wherein A comprises an amino acid sequence derived from a histone protein selected from the group consisting of histone HI, H2A, H2AX, H2B, H3 and H4,
- (b) performing a ChIP assay, thereby processing, in parallel to the genomic material of interest of (a), the one or more polypeptide-oligonucleotide conjugates of (a),
- (c) obtaining a processed sample of the polypeptide-oligonucleotide conjugate and the genomic material of interest processed in (b), and
- (d) analyzing the samples obtained in (c), thereby obtaining a value and/or signal for each of the analyzed processed samples, and
- (e) validating the ChIP assay result based on the value and/or signal obtained in (d).
4 more elements
A method of normalizing chromatin immunoprecipitation (ChIP) assay data, the method comprising:
- (a) obtaining an input sample of the genomic material of interest and of one or more of polypeptide-oligonucleotide conjugate of the formula:
- A-L-N,
- wherein A is a polypeptide comprising 5 amino acids, L is a linker, and N is an oligonucleotide comprising 10 nucleotides, wherein the sequence of the 10 nucleotides of N uniquely identifies an amino acid sequence and/or amino acid modification of A wherein A comprises an amino acid sequence derived from a histone protein selected from the group consisting of histone HI, H2A, H2AX, H2B, H3 and H4,
- (b) performing a ChIP assay, thereby processing, in parallel to the genomic material of interest in (a), the one or more polypeptide-oligonucleotide conjugates of (a),
- (c) obtaining a processed sample of the polypeptide-oligonucleotide conjugate and the genomic material of interest processed in (b),
- (d) analyzing the samples obtained in (c), thereby obtaining a value and/or signal for each of the analyzed processed samples, and
- (e) normalizing the values and/or signals obtained in (d) for each of the processed samples using the values obtained in (d) for each of the input samples.
4 more elements
Dependencies are inferred from the claim text.