title: Improved precision of proteomic measurements in immunoprecipitation based purifications using relative quantitation.
abstract: Mass spectrometry coupled immunoprecipitation (MS-IP) studies are useful in identifying and quantitating potential binding partners of a target protein. However, they are often conducted without appropriate loading controls. Western blots are often used to analyze loading controls, yet there are limitations to their usefulness as analytical tools. One remedy for this is the use of selected reaction monitoring (SRM), where the areas under the curve (AUCs) of peptides from a protein of interest can be normalized to those from the constant regions of the immunoglobulins used for the IP. Using this normalization method, significant changes in relative peptide abundance were observed between samples when there appeared to be an unequal load based on immunoglobulin peptide abundance.
authors.affiliation: Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, United States.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Rogstad
authors.forename: Tatiana
authors.initials: T
authors.lastname: Sorkina
authors.forename: Alexander
authors.initials: A
authors.lastname: Sorkin
authors.forename: Christine C
authors.initials: CC
authors.lastname: Wu
mesh_terms: D000818:Animals; D001011:Aorta / Q000737:chemistry / Q000166:cytology; D002460:Cell Line; D042783:Endothelial Cells / Q000737:chemistry / Q000166:cytology; D007136:Immunoglobulins / Q000032:analysis*; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D010455:Peptides / Q000032:analysis*; D040901:Proteomics; D013552:Swine
chemical_list: D007136:Immunoglobulins; D010455:Peptides